High affinity TCR proteins and methods

T cell receptors (TCRS) that have higher affinity for a ligand than wild type TCRs are provided. These high affinity TCRs are formed by mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants of the T cell receptor protein coding sequence; transforming the T cell receptor mutant coding sequence into yeast cells; inducing expression of the T cell receptor mutant coding sequence on the surface of yeast cells; and selecting those cells expressing T cell receptor mutants that have higher affinity for the peptide/MHC ligand than the wild type T cell receptor protein. The high affinity TCRs can be used in place of an antibody or single chain antibody.

System and method for specifying access to resources in a mobile code system

Mobile code, such as an applet, is permitted to create a network connection with a content server on a network, without restricting the applet only to connections from the computer from which it was downloaded. This is achieved in accordance with the principles of the present invention by using network restriction software in the execution engine or runtime system under which the applet executes. When the applet attempts to create a network connection to a content server, the network restriction software checks a name file on the content server for the presence of an entry whose name corresponds to the name of the computer from which the applet was downloaded. If such an entry is present, then the network restriction software permits the network connection between the applet and the content server to be created. If not, the applet may not create a network connection with the content server.

System and methods for securely permitting mobile code to access resources over a network

A system and methods are disclosed that permit mobile code, such as an applet, to create a network connection with a content server on a network, without exposing the client computer that is running the applet, or other computers with which the client computer may communicate, to a DNS spoofing attack. This is achieved in accordance with the principles of the present invention by using network restriction software in the execution engine or runtime system under which the applet executes. When the applet attempts to create a network connection to a content server, the network restriction software checks a “name directory” on the content server for the presence of an entry whose name corresponds to the name of the computer from which the applet was downloaded. If such an entry is present, then the network restriction software permits the network connection between the applet and the content server to be created. If not, the applet may not create a network connection with the content server. Additionally, address checks may be applied to assist in preventing DNS spoofing attacks from succeeding.

Using shifted syngas to regenerate SCR type catalyst

The present invention integrates a gasification unit into a catalyst/absorber process for removing pollutants from the combustion product of a gas turbine. A small slipstream of syngas from the gasification unit is cleaned in an acid gas removal unit to remove H 2 S. The syngas is then processed in a shift unit where the carbon monoxide and any COS present in the syngas are converted into hydrogen and carbon dioxide. The shifted syngas, still containing trace amounts of H 2 S, is then processed in a zinc oxide bed, where the trace H 2 S is removed. The resultant stream is hydrogen and carbon dioxide rich, making it ideal for use in regenerating the catalyst/absorber system.

System and methods for providing compatibility across multiple versions of a software system

A system and methods that provide compatibility across multiple versions of a software system, such as an execution engine or run-time system, are disclosed. The software system is structured so that it includes a master module, and one or more helper modules, each helper module being capable of processing applets (i.e., programs or content) that require a particular version of the software system. Each time an applet is to be processed by the software system, the master module determines which helper module should be used to process the applet, and starts the selected helper module if necessary. Compatibility with additional versions can be provided by providing additional helper modules. Because numerous helper modules may be executed simultaneously, applets that require different versions of the software system may be processed simultaneously.

Generator set tank and enclosure with adjustable mounting system

The present invention is a generator set tank and enclosure that includes a number of adjustable generator support assemblies that enable the tank and enclosure to be configured for use with generators of various sizes. The support assemblies include mounting channels configured to be secured to the tank in a variety of locations, and each channel includes a number of support platforms therein which can be adjustably positioned on the channels to further adjust the configuration of the support assemblies. Also, the tank and enclosure are configured with multiple fittings, attachments, and access points in order to enable both the tank and enclosure to be fully customizable for use with generators of various sizes.

Mutated class II major histocompatibility proteins

Provided are methods for directed mutagenesis and selection of MHC Class II proteins and single chain proteins with improved conformational stability and/or binding affinity for at least one cognate ligand. The improved proteins are useful for identification of T cells that are specific for the MHC Class II proteins and for treatment of disorders including autoimmune diseases, with the disorder determining the choice of the particular improved protein, peptide-protein complex or other ligand protein complex.

System and method supporting plural option data structures

To support values of properties, a class includes fields to support values in preallocated memory space and with an option data structure which supports, in instances of the class, references to option values without preallocation of memory space. The field and option values are accessed in an instance object of the class using expressions of the same syntactic form. During compilation, the compiler checks the type of an option value against a type description within the option data structure. If a value has not been set for an instance object, a get operation results in getting of the default value for the class. Different classes may support different forms of data structures such as a linked list or a hash table. During compilation, a method call to an object is encoded without regard to the form of the option data structure. When an option value is changed, a change handler identified by an option binding of the data structure is processed. That option binding may be located by first ...

System and method supporting property value as options

To support values of properties, a class includes fields to support values in preallocated memory space and with an option data structure which supports, in instances of the class, references to option values without preallocation of memory space. The field and option values are accessed in an instance object of the class using expressions of the same syntactic form. During compilation, the compiler checks the type of an option value against a type description within the option data structure. If a value has not been set for an instance object, a get operation results in getting of the default value for the class. Different classes may support different forms of data structures such as a linked list or a hash table. During compilation, a method call to an object is encoded without regard to the form of the option data structure. When an option value is changed, a change handler identified by an option binding of the data structure is processed. That option binding may be located by first ...

High affinity TCR proteins and methods

T cell receptors (TCRs) that have higher affinity for a ligand than wild type TCRs are provided. These high affinity TCRs are formed by mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants of the T cell receptor protein coding sequence; transforming the T cell receptor mutant coding sequence into yeast cells; inducing expression of the T cell receptor mutant coding sequence on the surface of yeast cells; and selecting those cells expressing T cell receptor mutants that have higher affinity for the peptide/MHC ligand than the wild type T cell receptor protein. The high affinity TCRs can be used in place of an antibody or single chain antibody.

Multiple source alignment sensor with improved optics

Features of the present invention provide an optical layout that can accommodate the relatively strict enclosure requirements for compact component alignment sensor. Specifically, aspects of the present invention provide a single optical component that reduces the degree of divergence, and preferably substantially collimates light from the plurality of divergent light sources prior to entering the component zone. In this regard, part count is kept low and the physical size of the optical train itself is relatively small.

Generator set tank and enclosure with adjustable mounting system

The present invention is a generator set tank and enclosure that includes a number of adjustable generator support assemblies that enable the tank and enclosure to be configured for use with generators of various sizes. The support assemblies include mounting channels configured to be secured to the tank in a variety of locations, and each channel includes a number of support platforms therein which can be adjustably positioned on the channels to further adjust the configuration of the support assemblies. Also, the tank and enclosure are configured with multiple fittings, attachments, and access points in order to enable both the tank and enclosure to be fully customizable for use with generators of various sizes.

Lazy compilation of template-generated classes in dynamic compilation execution environments

Template-generated classes in program code are compiled efficiently through a process of lazy compilation resulting in improved compilation times. Lazy compilation includes the generation of objects representing a class template and a template-generated class as well as the selective compilation of class methods that are invoked in the program code. Code sharing is a further enhancement for increasing compilation speed by providing a system and method for sharing executable object code for compatible methods among different classes generated from the same class template.

Lazy compilation of template-generated classes in dynamic compilation execution environments

Template-generated classes in program code are compiled efficiently through a process of lazy compilation resulting in improved compilation times. Lazy compilation includes the generation of objects representing a class template and a template-generated class as well as the selective compilation of class methods that are invoked in the program code. Code sharing is a further enhancement for increasing compilation speed by providing a system and method for sharing executable object code for compatible methods among different classes generated from the same class template.

System and method supporting type checking of options

To support values of properties, a class includes fields to support values in preallocated memory space and with an option data structure which supports, in instances of the class, references to option values without preallocation of memory space. The field and option values are accessed in an instance object of the class using expressions of the same syntactic form. During compilation, the compiler checks the type of an option value against a type description within the option data structure. If a value has not been set for an instance object, a get operation results in getting of the default value for the class. Different classes may support different forms of data structures such as a linked list or a hash table. During compilation, a method call to an object is encoded without regard to the form of the option data structure. When an option value is changed, a change handler identified by an option binding of the data structure is processed. That option binding may be located by first ...

URL system and method for licensing content

A licensing system and method for deploying digital content is provided. A client system can download the digital content from the server and, if a valid license for the content is also found on the server, software may be permitted to run digital content. The license is stored in a location that is separate from the content. The license can be located using the URL address of the content. The system can locate the license on the server and determine whether the license covers the content. Content that is covered by the license can be processed and rendered. The content can be any form of digital media, such as a computer application.

Yeast cell surface display of proteins and uses thereof

The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. The outer wall of each yeast cell can display approximately 10 4 protein agglutinins. The native agglutinins serve as specific adhesion contacts to fuse yeast cells of opposite mating type during mating. In effect, yeast has evolved a platform for protein-protein binding without steric hindrance from cell wall components. As one embodiment, attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. As another embodiment, T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of alteting T cell receptor binding affinity and specificity by library screening.

Mutated class I major histocompatibility proteins and complexes

Provided herein are combinatorial libraries containing chimeric Major Histocompatibility Comples (MHC) Class I proteins displayed on the surfaces of recombinant yeast cells. Members of the libraries, especially where those libraries have been mutagenized either with error-prone Polymerase Chain Reaction or with site-directed oligonucleotide mutagenesis, are improved in conformation stability or in binding to a target, e.g., a peptide or other ligand as compared with the stability or binding affinity of a wild type MHC Class I chimeric protein. The improved mutant chimeric proteins can be selected by various means, including fluorescence activated cell sorting with a fluorescent ligand bound to the surfaces of the yeast cells displaying the improved mutant chimeric protein.

Safe I/O through use of opaque I/O objects

Opaque I/O objects are described which can be used in a variety of mobile code systems to permit unprivileged applets to perform a wide variety of I/O operations in a safe manner. Such opaque objects permit limited I/O without requiring the user of an applet to confer trust or privilege on the applet, and without exposing the user to a risk of his data being destroyed, compromised, or stolen by malicious applets.

System and method supporting nonlocal values

To support values of properties, a class includes fields to support values in preallocated memory space and with an option data structure which supports, in instances of the class, references to option values without preallocation of memory space. The field and option values are accessed in an instance object of the class using expressions of the same syntactic form. During compilation, the compiler checks the type of an option value against a type description within the option data structure. If a value has not been set for an instance object, a get operation results in getting of the default value for the class. Different classes may support different forms of data structures such as a linked list or a hash table. During compilation, a method call to an object is encoded without regard to the form of the option data structure. When an option value is changed, a change handler identified by an option binding of the data structure is processed. That option binding may be located by first searching a mapping data structure for a previously computed mapping to the option binding or by computing the mapping to the option binding. An option value may be set in an option data structure from an initialization expression which includes the name of the option value and, as an argument, the option value. Nonlocal option values may be applied to plural objects in a nonlocal option hierarchy such as a graphical hierarchy.

Automated system with improved height sensing

An automated system includes improved height sensing. In one aspect, a on-head camera performs the dual functions of fiducial imaging and height sensing using an auxiliary off-axis light source and triangulation. In another aspect, an on-head height sensor is positioned to measure height at a location that is not beneath any nozzles. The sensor provides height information at a plurality of locations over the board, and a height map of the board is created. In yet another aspect of the invention, the above features are combined to provide an on-head camera that images fiducials and measures height at a plurality of locations such that a height map is created.

Lazy compilation of template-generated classes in dynamic compilation execution environments

Template-generated classes in program code are compiled efficiently through a process of lazy compilation resulting in improved compilation times. Lazy compilation includes the generation of objects representing a class template and a template-generated class as well as the selective compilation of class methods that are invoked in the program code. Code sharing is a further enhancement for increasing compilation speed by providing a system and method for sharing executable object code for compatible methods among different classes generated from the same class template.

System and method supporting mapping of option bindings

To support values of properties, a class includes fields to support values in preallocated memory space and with an option data structure which supports, in instances of the class, references to option values without preallocation of memory space. The field and option values are accessed in an instance object of the class using expressions of the same syntactic form. During compilation, the compiler checks the type of an option value against a type description within the option data structure. If a value has not been set for an instance object, a get operation results in getting of the default value for the class. Different classes may support different forms of data structures such as a linked list or a hash table. During compilation, a method call to an object is encoded without regard to the form of the option data structure. When an option value is changed, a change handler identified by an option binding of the data structure is processed. That option binding may be located by first ...

Phase profilometry system with telecentric projector

... an optical system for computing a height of a target on a surface includes a light projector for projecting light. The light passes through a patterned reticle and a projector lens so as to illuminate the target with an image of the pattern. The light is projected telecentrically between the reticle and the projector lens, and a camera is positioned along a receive optical path. The camera receives an image of the target through a receive lens. The target and the pattern move at least three times with respect to each other, and the camera acquires an image of the object at each of at least three positions.

Human Single-Chain T Cell Receptors

A soluble human single-chain T cell receptor (TCR) having the structure: Vα2-L-Vβ or Vβ-L-Vα2, wherein L is a linker peptide that links Vβ with Vα, Vβ is a TCR variable β region, and Vα2 is a TCR variable α region of the family 2 is provided. The provided scTCR is useful for many purposes, including the treatment of cancer, viral diseases and autoimmune diseases. 1. A soluble human single-chain T cell receptor (TCR) having the structure: Vα2-L-Vβ or Vβ-L-Vα2 , wherein L is a linker peptide that links Vβ with Vα , Vβ is a TCR variable β region , and Vα2 is a TCR variable a region of the family 2.2. The TCR of claim 1 , wherein the Vα2 region is subfamily Vα2.1.3. The TCR of claim 1 , wherein the Vα2 region contains a mutation at position 49.4. The TCR of claim 3 , wherein the mutation is PheSer.5. The TCR of claim 1 , further comprising one or more mutations in CDR2β or CDR3α.6. The TCR of claim 1 , wherein the linker peptide contains more than 5 lysine residues.7. The TCR of claim 1 , wherein the linker peptide contains between 10 and 30 amino acids.8. The TCR of claim 1 , wherein the linker peptide is GSADDAKKDAAKKDGKS [SEQ ID NO: 4].9. The TCR of which does not contain a constant region.10. The TCR of wherein the TCR binds specifically to a ligand with an equilibrium binding constant Kof between about 10M and 10M.11. The TCR of claim 10 , wherein the ligand is a peptide/MHC ligand.12. The TCR of claim 10 , wherein the ligand includes a peptide known as SL9 from the HIV Gag protein.13. The TCR of claim 10 , wherein the ligand includes a peptide known as MART-1 expressed by melanoma.14. The TCR of that has the sequence of the Vα and Vβ regions from the scTCR known as 868-Z11.15. The TCR of which is functionally equivalent to the sequence of the Vα and Vβ regions from the scTCR known as 868-Z11.16. The TCR of any of the preceding claims claim 10 , further comprising a biologically active group.17. The TCR of claim 16 , wherein the biologically active group is ...

Yeast Cell Surface Display of Proteins and Uses Thereof

The present invention provides a genetic method of tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. As one embodiment, attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. As another embodiment, T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of altering T cell receptor binding affinity and specificity by library screening. 1126-. (canceled)127. A method of producing a yeast cell displayed variant ligand binding protein with enhanced binding properties relative to a wild-type of said ligand binding protein , the method comprising:isolating a gene encoding said wild-type binding protein;creating a library of mutated proteins by randomly mutating said wild-type protein; {'br': None, '5′-GAL1-10-aAga2p-mutated polypeptide;'}, 'incorporating each said mutated protein into respective expression cassettes, each having the structure'}incorporating each said expression cassette into a respective vector;transforming yeast cells with said cassette-containing vectors to yield a multiplicity of transformed yeast cells;expressing said cassettes in said transformed yeast cells, whereby said ligand binding protein is displayed on the surface of each said yeast cell, said displayed ligand binding protein containing one of said mutated binding;labeling the displayed proteins on said yeast cells by binding a specific label to said displayed proteins;employing flow cytometry to sort said yeast cells according to their labeling characteristics;determining the surface expression level of said ligand binding protein in said ...

Cancer treatment with 237 car-t cell based therapeutics recognizing the tn epitope

The disclosure provides Tn epitope-specific chimeric antigen receptors and scFvs, including soluble scFvs and multimeric scFvs, as well as methods of identifying cancer subjects and cancer subject sub-populations amenable to anti-Tn immunotherapy and methods of treating cancer.

ENGINEERED HIGH-AFFINITY HUMAN T CELL RECEPTORS

T cell receptors (TCRs) that have higher affinity for the Survivin antigen are provided. The high affinity TCRs were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the engineered TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting. 1. A modified T cell receptor , or antigen-binding fragment thereof , comprising a Vα and a Vβ derived from a wild type T cell receptor , wherein the Vα or the Vβ , or both , comprise a mutation in one or more complementarity determining regions (CDRs) relative to the wild type T cell receptor , wherein the modified T cell receptor binds to a complex of the peptide Survivin and the HLA-A2 molecule with a nanomolar or higher affinity (Kvalue of less than 10M).2. The modified T cell receptor of claim 1 , wherein the modified T cell receptor comprises the Vα amino acid sequence set forth in SEQ ID NO:1.3. The modified T cell receptor of claim 1 , wherein the modified T cell receptor comprises the Vα amino acid sequence set forth in SEQ ID NO:2.4. The modified T cell receptor of claim 1 , wherein the modified T cell receptor comprises the single-chain T cell receptor with the amino acid sequence set forth in SEQ ID NO:3.5. The modified T cell receptor of claim 1 , wherein the modified T cell receptor comprises the single-chain T cell receptor with the amino acid sequence set forth in SEQ ID NO:4.6. The modified T cell receptor of claim 1 , comprising a modified Vα comprising an amino acid sequence having at least 80% identity to the Vα amino acid sequence set forth in SEQ ID NO:1.7. The modified T cell receptor of claim 1 , comprising a modified Vα comprising an amino acid sequence having at least 80% identity to the Vα amino acid sequence set forth in SEQ ID NO:2.8. The modified T cell receptor of ...

ENGINEERING T CELL RECEPTORS

The use of model T cell receptors (TCRs) as scaffolds for in vitro engineering of novel specificities is provided. TCRs with de novo binding to a specific peptide-major histocompatibility complex (MHC) product can be isolated by: 1) mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants (a library), 2) selection of the library of TCR mutants with the specific peptide-MHC, using a process of directed evolution and a “display” methodology (e.g., yeast, phage, mammalian cell) and the peptide-MHC ligand. The process can be repeated to identify TCR variants with improved affinity for the selecting peptide-MHC ligand. 1. A modified T cell receptor , or antigen binding fragment thereof , comprising a Vα and a Vβ derived from a wild type T cell receptor , wherein the Vα , the Vβ , or both , comprise a mutation in one or more complementarity determining regions (CDRs) relative to the wild type T cell receptor , wherein the modified T cell receptor binds to a non-cognate peptide-MHC not bound by the wild type T cell receptor.2. The modified T cell receptor of claim 1 , wherein the wild type T cell receptor comprises the Vα amino acid sequence set forth in SEQ ID NO:1 and the Vβ amino acid sequence set forth in SEQ ID NO:2.3. The modified T cell receptor of claim 2 , comprising a modified Vα comprising an amino acid sequence having at least 80% identity to the Vα amino acid sequence set forth in SEQ ID NO:1 and a modified Vβ comprising an amino acid sequence having at least 80% identity to the Vβ amino acid sequence set forth in SEQ ID NO:2 claim 2 , wherein the modified T cell receptor does not bind to the cognate peptide-MHC bound by the wild type T cell receptor.4. The modified T cell receptor of claim 2 , wherein the modified T cell receptor comprises an amino acid substitution at one or more of CDR1α 31 claim 2 , CDR3α 98 claim 2 , CDR3β 99 claim 2 , CDR3α 97 claim 2 , CDR3β 102 claim 2 , CDR3α 99 claim 2 , CDR3β 100 claim 2 ...

High Affinity TCR Proteins and Methods

T cell receptors (TCRs) that have higher affinity for a ligand than wild type TCRs are provided. These high affinity TCRs are formed by mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants of the T cell receptor protein coding sequence; transforming the T cell receptor mutant coding sequence into yeast cells; inducing expression of the T cell receptor mutant coding sequence on the surface of yeast cells; and selecting those cells expressing T cell receptor mutants that have higher affinity for the peptide/MHC ligand than the wild type T cell receptor protein. The high affinity TCRs can be used in place of an antibody or single chain antibody. 1. A method for cloning the gene for a high affinity TCR mutant into a system that allows expression of the mutant on the surface of T cells comprising:mutating TCRs to create high affinity TCR mutants;cloning said TCR mutants into a vector;transfecting the vector into T cells;expressing the high affinity TCR mutant on the surface of T cells.2. The method of claim 1 , further comprising:selecting those T cells that are activated by a peptide/MHC ligand more than the wild type.3. The method of claim 1 , wherein the transfected/infected T cells are used for recognition of selected peptide-bearing MHC cells.4. T cells made by the method of .5. A method for cloning the gene for a high affinity TCR mutant into a system that allows expression of the mutant on the surface of T cells comprising the steps of:{'sup': 7', '−1, 'mutating TCRs to create high affinity TCR mutants which exhibit a dissociation constant for their cognate ligand of at least about 10M;'}cloning said TCR mutants into a vector;transfecting the vector into T cells; andexpressing the high affinity TCR mutant on the surface of T cells.6. The method of claim 5 , wherein the transfected T cells are used for recognition of selected peptide-bearing MHC cells.7. The method of wherein the high affinity TCR mutants carry one ...

ENGINEERED HIGH-AFFINITY HUMAN T CELL RECEPTORS

T cell receptors (TCRs) that have higher affinity for the Survivin antigen are provided. The high affinity TCRs were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the engineered TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting. 1. A modified T cell receptor , or antigen-binding fragment thereof , comprising a Vα and a Vβ derived from a wild type T cell receptor , wherein the Vα or the Vβ , or both , comprise a mutation in one or more complementarity determining regions (CDRs) relative to the wild type T cell receptor , wherein the modified T cell receptor binds to a complex of the peptide Survivin and the HLA-A2 molecule with a nanomolar or higher affinity (Kvalue of less than 10M).2. The modified T cell receptor of claim 1 , wherein the modified T cell receptor comprises the Vα amino acid sequence set forth in SEQ ID NO:1.3. The modified T cell receptor of claim 1 , wherein the modified T cell receptor comprises the Vα amino acid sequence set forth in SEQ ID NO:2.4. The modified T cell receptor of claim 1 , wherein the modified T cell receptor comprises the single-chain T cell receptor with the amino acid sequence set forth in SEQ ID NO:3.5. The modified T cell receptor of claim 1 , wherein the modified T cell receptor comprises the single-chain T cell receptor with the amino acid sequence set forth in SEQ ID NO:4.6. The modified T cell receptor of claim 1 , comprising a modified Vα comprising an amino acid sequence having at least 80% identity to the Vα amino acid sequence set forth in SEQ ID NO:1.7. The modified T cell receptor of claim 1 , comprising a modified Vα comprising an amino acid sequence having at least 80% identity to the Vα amino acid sequence set forth in SEQ ID NO:2.8. The modified T cell receptor of ...

ENGINEERED HIGH-AFFINITY HUMAN T CELL RECEPTORS

T cell receptors (TCRs) that have specificity for the WT1 antigen are provided. The TCRs include higher affinity TCRs that were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting. 1. A T cell receptor , or antigen-binding fragment thereof , comprising a Vα and a Vβ derived from a T cell clone , wherein the T cell receptor binds to a complex of the peptide WT1 and the HLA-A2 molecule , and wherein the T cell receptor comprises the Vβ amino acid sequence set forth in SEQ ID NO:1 and the Vα amino acid sequence set forth in SEQ ID NO:2.2. A host cell that expresses the T cell receptor of .3. The host cell of claim 2 , wherein the cell is a human T cell.4. A modified T cell receptor claim 2 , or antigen-binding fragment thereof claim 2 , comprising a Va and a Vβ derived from a wild type T cell receptor claim 2 , wherein the Vα or the Vβ claim 2 , or both claim 2 , comprise a mutation in one or more complementarity determining regions (CDRs) relative to the wild type T cell receptor claim 2 , wherein the modified T cell receptor binds to a complex of the peptide WT1 and the HLA-A2 molecule with higher affinity than the wild type T cell receptor.5. The modified T cell receptor of claim 4 , wherein the modified T cell receptor comprises the Vβ amino acid sequence set forth in SEQ ID NO:3 and the Vα amino acid sequence set forth in SEQ ID NO:4.6. The modified T cell receptor of claim 4 , wherein the modified T cell receptor comprises the single-chain T cell receptor with the amino acid sequence set forth in SEQ ID NO:5.7. The modified T cell receptor of claim 4 , comprising a modified Vα comprising an amino acid sequence having at least 80% identity to the Vα amino acid sequence ...

Engineering t cell receptors

The use of model T cell receptors (TCRs) as scaffolds for in vitro engineering of novel specificities is provided. TCRs with de novo binding to a specific peptide-major histocompatibility complex (MHC) product can be isolated by: 1) mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants (a library), 2) selection of the library of TCR mutants with the specific peptide-MHC, using a process of directed evolution and a “display” methodology (e.g., yeast, phage, mammalian cell) and the peptide-MHC ligand. The process can be repeated to identify TCR variants with improved affinity for the selecting peptide-MHC ligand.

Conjugates of folate anti-effector cell antibodies

The present invention provides a process of targeting folate-receptor-positive tumor cells for lysis by binding a conjugate of folate and an anti-T-cell-receptor antibody or an anti-Fc receptor antibody to those cells. A process of lysing folate-receptor-positive tumor cells comprising exposing the cells to a folate/anti-T-cell-receptor antibody in the presence of a population of T-cells is also provided. A process of lysing folate-receptor-positive tumor cells comprising exposing the cells to a folate/anti-Fc receptor antibody in the presence of a population of natural killer cells, monocytes, or macrophages is also provided. Still further, the present invention provides a conjugate of folate to an anti-T-cell-receptor antibody or an anti-Fc receptor antibody.

Yeast cell surface display of proteins and uses thereof

The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. The outer wall of each yeast cell can display approximately 10 4 protein agglutinins. The native agglutinins serve as specific adhesion contacts to fuse yeast cells of opposite mating type during mating. In effect, yeast has evolved a platform for protein-protein binding without steric hindrance from cell wall components. As one embodiment, attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. As another embodiment, T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of altering T cell receptor binding affinity and specificity by library screening.

High affinity TCR proteins and methods

T cell receptors (TCRs) that have higher affinity for a ligand than wild type TCRs are provided. These high affinity TCRs are formed by mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants of the T cell receptor protein coding sequence; transforming the T cell receptor mutant coding sequence into yeast cells; inducing expression of the T cell receptor mutant coding sequence on the surface of yeast cells; and selecting those cells expressing T cell receptor mutants that have higher affinity for the peptide/MHC ligand than the wild type T cell receptor protein. The high affinity TCRs can be used in place of an antibody or single chain antibody.

Yeast cell surface display of proteins and uses thereof

The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. The outer wall of each yeast cell can display approximately 10 4 protein agglutinins. The native agglutinins serve as specific adhesion contacts to fuse yeast cells of opposite mating type during mating. In effect, yeast has evolved a platform for protein-protein binding without steric hindrance from cell wall components. As one embodiment, attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. As another embodiment, T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of altering T cell receptor binding affinity and specificity by library screening.

Yeast cell surface display of proteins and uses thereof

The present invention provides a genetic method of tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. As one embodiment, attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. As another embodiment, T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of altering T cell receptor binding affinity and specificity by library screening.

Yeast cell surface display of proteins and uses thereof

The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. Attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of altering T cell receptor binding affinity and specificity by library screening. As another embodiment, the selection method identifies proteins displayed at higher levels as proteins that are secreted at higher efficiency and that are more stable.

Yeast cell surface display of proteins and uses thereof

The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. The outer wall of each yeast cell can display approximately 10 4 protein agglutinins. The native agglutinins serve as specific adhesion contacts to fuse yeast cells of opposite mating type during mating. If effect, yeast has evolved a platform for protein-protein binding without steric hindrance from cell wall components. As one embodiment, attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. As another embodiment, T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of altering T cell receptor binding affinity and specificity by library screening.

Yeast cell surface display of proteins and uses thereof

The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. The outer wall of each yeast cell can display approximately 10 4 protein agglutinins. The native agglutinins serve as specific adhesion contacts to fuse yeast cells of opposite mating type during mating. In effect, yeast has evolved a platform for protein-protein binding without steric hindrance, from cell wall components. As one embodiment, attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. As another embodiment, T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of altering T cell receptor binding affinity and specificity by library screening.

Mutated class ii major histocompatibility proteins

Provided are methods for directed mutagenesis and selection of MHC Class II proteins and single chain proteins with improved conformational stability and/or binding affinity for at least one cognate ligand. THe improved proteins are useful for identificaiton of T cells that are specific for the MHC Class II proteins and for treatment of disorders inlcuding autoimmune diseases, with the disorder determining the choice of the particular improved protein, peptide-protein complex or other ligand protein complex.

High affinity tcr proteins and methods

T cell receptors (TCRs) that have higher affinity for ligand than wild type TCRs are provided. These high affinity TCRs are formed by mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants of the T cell receptor protein coding sequence; transforming the T cell receptor mutant coding sequence into yeast cells; inducing expression of the T cell receptor mutant coding sequence on the surface of yeast cells; and selecting those cells expressing T cell receptor mutants that have higher affinity for the peptide/MHC ligand than the wild type T cell receptor protein. The high affinity TCRs can be used in place of an antibody or single chain antibody.

Engineering T-cell receptors

Il:\aar\Interwovn\NRPortbl\DCC\AAR278367_l.docx-I1/01/2018 The use of model T cell receptors (TCRs) as scaffolds for in vitro engineering of novel specificities is provided. TCRs with de novo binding to a specific peptide major histocompatibility complex (MHC) product can be isolated by: 1) mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants (a library), 2) selection of the library of TCR mutants with the specific peptide MHC, using a process of directed evolution and a "display" methodology (e.g., yeast, phage, mammalian cell) and the peptide-MHC ligand. The process can be repeated to identify TCR variants with improved affinity for the selecting peptide-MHC ligand.

genetically modified high affinity human t cell receptors

Mutated class ii major histocompatibility proteins

System and method supporting type checking of options

To support values of properties, a class includes fields to support values in preallocated memory space and with an option data structure which supports, in instances of the class, references to option values without preallocation of memory space. The field and option values are accessed in an instance object of the class using expressions of the same syntactic form. During compilation, the compiler checks the type of an option value against a type description within the option data structure. Different classes may support different forms of data structures such as a linked list or a hash table. During compilation, a method call to an object is encoded without regard to the form of the option data structure. An option value may be set in an option data structure from an initialization expression which includes the name of the option value and, as an argument, the option value.

Engineered high-affinity human t cell receptors

Using shifted syngas to regenerate SCR type catalyst

Engineered high-affinity human t cell receptors

Genes encoding the 5h7 antibody and methods for conferring programmed cell death properties to cells

The present invention relates to isolated and purified polynucleotides encoding for the light and heavy variable regions of a 5H7 antibody and methods for using these genes to confer programmed cell death properties to a cell.

High affinity tcr proteins and methods

T cell receptors (TCRs) that have higher affinity for ligand than wild type TCRs are provided. These high affinity TCRs are formed by mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants of the T cell receptor protein coding sequence; transforming the T cell receptor mutant coding sequence into yeast cells; inducing expression of the T cell receptor mutant coding sequence on the surface of yeast cells; and selecting those cells expressing T cell receptor mutants that have higher affinity for the peptide/MHC ligand than the wild type T cell receptor protein. The high affinity TCRs can be used in place of an antibody or single chain antibody.

Mutated class i major histocompatibility proteins and complexes

Human single-chain t cell receptors

A soluble human single-chain T cell receptor (TCR) having the structure: Va2-L-Vß or Vß-L-Va2, wherein L is a linker peptide that links Vß with Va, Vß is a TCR variable ß region, and Va2 is a TCR variable a region of the family 2 is provided. The provided scTCR is useful for many purposes, including the treatment of cancer, viral diseases and autoimmune diseases.

Treatment of diseases caused by bacterial exotoxins

Provided are high affinity T cell receptor variable regions that are useful for treating diseases caused by superantigens including atopic dermatitis, pneumonia and delayed wound healing. The variable regions contain mutants that result in high affinity binding to the superantigen.

Yeast cell surface display of proteins and uses thereof

【課題】野生型タンパク質の表現型特性と比較して増強した表現型特性を有するタンパク質を選択するための方法の提供。 【解決手段】酵母細胞壁タンパク質に融合される、試験されるタンパク質を発現するベクターを用いて酵母細胞を形質転換する工程であって、ここで変異誘発を使用し、該試験されるタンパク質の変異体の変化に富む集団を生成する工程と、該酵母細胞を第１の標識で標識する工程であって、ここで該第１の標識が該試験されるタンパク質を発現する酵母と結合し及び該試験されるタンパク質を発現しない酵母とは結合しない工程と、該第１の標識が結合する該酵母細胞を単離する工程と酵母により発現される該変異タンパク質の該特性を分析し、そして該野生型タンパク質の特性と比較する工程であって、ここで野生型タンパク質に対して増強した特性を提示する変異タンパク質を有する酵母細胞が選択される工程と、を含む、方法。 【選択図】なし

High affinity TCR proteins and methods

T cell receptors (TCRs) that have higher affinity for ligand than wild type TCRs are provided. These high affinity TCRs are formed by mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants of the T cell receptor protein coding sequence; transforming the T cell receptor mutant coding sequence into yeast cells; inducing expression of the T cell receptor mutant coding sequence on the surface of yeast cells; and selecting those cells expressing T cell receptor mutants that have higher affinity for the peptide/MHC ligand than the wild type T cell receptor protein. The high affinity TCRs can be used in place of an antibody or single chain antibody.

Yeast cell surface display of proteins and uses thereof

The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. Attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. T cell receptor mutants that are efficiently displayed on the yeast cell surface can be isolated by this method, providing a means of altering T cell receptor binding affinity and specificity by library screening. The selection method also identifies proteins with enhanced phenotypic characteristics, proteins that are displayed at higher levels, proteins that are secreted at higher efficiency and proteins that are more stable.

Engineered high-affinity human t cell receptors

T cell receptors (TCRs) that have specificity for the WT1 antigen are provided. The TCRs include higher affinity TCRs that were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting.

Engineering t-cell receptors

Disclosed are modified T cell reeceptors binding a SURV:HLA-A2 survivin antigen MHC complex. The disclosed receptors are derived from the Tax specific A6 TCR and comprise amino acid substitutions at positions 31, 32, 97, 98 and 99 on the alpha chain and positions 30 99, 100 and 102 on the beta chain. The use of model T cell receptors (TCRs) as scaffolds for in vitro engineering of novel specificities is provided. TCRs with de novo binding to a specific peptide-major histocompatibility complex (MHC) product can be isolated by: 1) mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants (a library), 2) selection of the library of TCR mutants with the specific peptide-MHC, using a process of directed evolution and a “display” methodology (e.g., yeast, phage, mammalian cell) and the peptide-MHC ligand. The process can be repeated to identify TCR variants with improved affinity for the selecting peptide-MHC ligand.

Allogeneic histocompatibility complexes as mediators of cell destruction

Methods are disclosed of producing and using Major Histocompatibility Complex (MHC) proteins, in native, truncated of fusion forms, complexed with MHC binding peptides and conjugated to targeting molecules, in order to target selected cells in a subject for T cell mediated destruction in vivo. The MHC proteins are chosen so to be allogeneic with respect to the host and the targeting molecules are chosen to selectively bind to the target cells. Pharmaceutical preparations including the targeted peptide-MHC complexes are also disclosed.

Yeast cell surface of proteins having enhanced properties

The present invention provides a method for selecting proteins with one or more enhanced phenotypic properties of surface expression level, stability, secretion levels and solubility relative to those of a wild-type of said protein. Yeast cells are transformed with a vector expressing a protein to be tested fused to a yeast cell wall protein. Mutagenesis is used to a generate a variegated population of mutants of the protein to be tested. The yeast cells are labeled with a first label which associates with yeast expressing the protein having one or more enhanced phenotypic properties of surface expression level, stability, secretion levels and solubility and does not associate with yeast which do not express the protein having enhanced phenotypic properties, and the transformed yeast cells with which the first label is associated are isolated. After analyzing and comparing the phenotypic properties of the mutant protein expressed by yeast with phenotypic properties of the wild-type protein, yeast cells exhibiting proteins with one or more enhanced phenotypic properties of surface expression level, stability, secretion levels and solubility over the wild-type protein are selected.

High affinity tcr proteins and methods

T cell receptors (TCRs) that have higher affinity for ligand than wild type TCRs are provided. These high affinity TCRs are formed by mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants of the T cell receptor protein coding sequence; transforming the T cell receptor mutant coding sequence into yeast cells; inducing expression of the T cell receptor mutant coding sequence on the surface of yeast cells; and selecting those cells expressing T cell receptor mutants that have higher affinity for the peptide/MHC ligand than the wild type T cell receptor protein. The high affinity TCRs can be used in place of an antibody or single chain antibody.

Process for making a targeted cell susceptible to lysis by cytotoxic t lymphocytes

A process whereby any cell can be made susceptible to lysis by cytotoxic T lymphocytes is disclosed. The process consists of binding or chemically linking an antibody, specific for portions of a T lymphocyte receptor shared by most cytotoxic T lymphocytes or a receptor-associated molecule, to the surface of the targeted cell. Recognition of or interaction with the reactive site of the antibody by the T lymphocyte results in lysis of the targeted cell. The antibody to the T lymphocyte receptor or associated molecule may be bound directly to the targeted cell using a crosslinking reagent or indirectly by means of a molecule such as an antibody, a hormone, or a carbohydrate which binds to particular molecules on the targeted cell's surface.

Engineered high-affinity human t cell receptors

System and method for specifying access to resources in a mobile code system

Mobile code, such as an applet, is permitted to create a network connection with a content server on a network, without restricting the applet only to connections from the computer from which it was downloaded. This is achieved in accordance with the principles of the present invention by using network restriction software in the execution engine or runtime system under which the applet executes. When the applet attempts to create a network connection to a content server, the network restriction software checks a name file on the content server for the presence of an entry whose name corresponds to the name of the computer from which the applet was downloaded. If such an entry is present, then the network restriction software permits the network connection between the applet and the content server to be created. If not, the applet may not create a network connection with the content server.

Multiple source alignment sensor with improved optics

Features of the present invention provide an optical layout that can accommodate the relatively strict enclosure requirements for compact component alignment sensor. Specifically, aspects of the present invention provide a single optical component (217) that reduces the degree of divergence, and preferably substantially collimates light from the plurality of divergent light sources (212, 214) prior to entering the component zone. In this regard, part count is kept low and the physical size of the optical train itself is relatively small.

Heterodimeric T lymphocyte receptor

Disclosed is a heterodimeric T lymphocyte receptor subunit. The subunit consists of a signal peptide, variable, joining, constant, transmembrane, and cytoplasmic regions. The structure, amino acid, and nucleotide sequence of the lymphocyte receptor subunit were determined using cDNA clones derived from a functional murine cytotoxic T lymphocyte clone. The genes corresponding to these cDNA are expressed and rearranged specifically in T cells and have significant sequence homologies to immunoglobulin V and C genes. T cell receptor subunits may be produced from the cDNA clones. The protein molecules may be further used for the production of T-cell clone specific antibodies.

Yeast cell surface display of T cell receptors

Engineering t-cell receptors

The use of model T cell receptors (TCRs) as scaffolds forin vitroengineering of novel specificities is provided. TCRs with de novo binding to a specific peptide-major histocompatibility complex (MHC) product can be isolated by: 1) mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants (a library), 2) selection of the library of TCR mutants with the specific peptide-MHC, using a process of directed evolution and a "display" methodology (e.g., yeast, phage, mammalian cell) and the peptide-MHC ligand. The process can be repeated to identify TCR variants with improved affinity for the selecting peptide-MHC ligand.

Engineered high-affinity human T cell receptors

T cell receptors (TCRs) that have higher affinity for the Survivin antigen are provided. The high affinity TCRs were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the engineered TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting.

Heterodimeric t lymphocyte receptor

Disclosed is a heterodimeric T lymphocyte receptor comprising an alpha and a beta subunit. Each subunit consists of a signal peptide, variable, joining, constant, transmembrane, and cytoplasmic regions. The two subunits are connected by a disulfide bond between cysteine residues located between the constant and transmembrane region. The structure, amino acid, and nucleotide sequence of the lymphocyte receptor were determined using cDNA clones derived from a functional murine cytotoxic T lympho­ cyte clone. The genes corresponding to these cDNA are expressed and rearranged specifically in T cells and have significant sequence homologies to immunoglobulin V and C genes. Both the T cell receptor protein and it subunits may be produced from the cDNA clones. The protein molecules may be further used for the production of T-cell clone specific antibodies.

Engineered high-affinity human T cell receptors

T cell receptors (TCRs) that have specificity for the WT1 antigen are provided. The TCRs include higher affinity TCRs that were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting.

Solder paste inspection system

A novel inspection system (10) for inspecting an article of manufacture, such as a printed circuit board (22), is disclosed, where the system (16) includes a strobed illuminator (38) adapted to project light through a reticle (41) so as to project a pattern of light onto an area of the printed circuit board. A board transport (16) responsively positions the board (22) to at least two distinct positions, where each position corresponding to a different phase of the projected light. Also included is a detector (18) adapted to acquire at least two images of the area, each image corresponding to one of the at least two different phases. An encoder (24) monitors the movement of the board (22) and outputs a position output (20), and a processor (14) connected to the encoder (24), the board transport (16), the illuminator (38) and the detector (18) controlledly energizes the illuminator (18) to expose the area as a function of the position output (20), the processor (14) co-siting the at least two images and constructing a height map image (4) with the co-sited images.

Multiple source alignment sensor with improved optics

Dark field illuminator with large working area

An illuminator is described which may be used with large inspection areas and which provides a dark field illumination pattern that is spatially uniform, illuminates from consistent angles, has high efficiency, and is smaller than existing solutions. A light pipe has a first end proximate an object to be illuminated and a second end opposite the first end and spaced from the first end. The light pipe also has at least one reflective sidewall. The first end of the light pipe includes an exit aperture and the second end has at least one opening to allow at least one image acquisition device to view the surface therethrough. At least one light source is configured to provide illumination in the light pipe. The object is illuminated by the first end of the light pipe by illumination at a selected elevation angle and substantially all azimuth angles.

Surface-bound antigen binding portions of antibodies that bind to ctla-4 and cd28 and uses therefor

The instant invention provides compositions for downmodulation or upmodulation of the immune response. For example, constructs comprising an exposed surface having attached to it i) an antigen-binding portion of an antibody that binds to a CTLA-4 or CD28 molecule that is expressed on a T cell of the subject, and ii) an MHC molecule selected from the group consisting of: a class II molecule that is syngeneic to the subject, a class I molecule that is syngeneic to the subject, and a class I molecule that is allogeneic to the subject are provided. Methods of using the molecules of the invention in downmodulation or upmodulation of the immune response are also provided. Also provided are methods of downmodulating an immune response in a subject comprising causing a cell of the subject to express an antigen-binding portion of an antibody that binds a CTLA-4 or CD28 molecule that is expressed on a T cell of the subject, such that the immune response in the subject is downmodulated. Methods of preparing an allogeneic cell for transplantation and methods of transplanting engineered allogeneic cells are also provided.

MODIFIED HIGH AFFINITY HUMAN T-CELLE RECEPTORS

Interface frames for threads

Multiple source alignment sensor with improved optics

Sensorsystem für die Berechnung von Anordnungsinformation über ein Bauteil in einem Bearbeitungsapparat für elektronische Bauteile, wobei der Apparat das Bauteil lösbar hält und geeignet ist, das Bauteil zu drehen, wobei das Sensorsystem aufweist: einen Sensor; mehrere divergente Lichtquellen in dem Sensor, die derart angeordnet sind, daß sie einen Bauteilbereich in dem Sensor beleuchten; einen Detektor, der relativ zu den Lichtquellen so angeordnet ist, daß das Bauteil, wenn das Bauteil zumindest teilweise in dem Bauteilbereich angeordnet ist, zumindest etwas von der Beleuchtung von mindestens einer der mehreren divergenten Lichtquellen abschirmt, um einen Schatten von zumindest einem Teil des Bauteils auf dem Detektor zu bilden, wobei der Detektor geeignet ist, während sich das Bauteil dreht, mehrere Detektorausgaben bereitzustellen; eine zwischen einen Bauteilbereich und die mehreren divergenten Lichtquellen eingefügte Optik, um die Divergenz des durch sie durchgehenden Lichts zu verringern; und eine Berechnungselektronik, welche die Detektorausgaben empfängt, um... A sensor system for computing placement information about a component in an electronic component processing apparatus, the apparatus releasably holding the component and adapted to rotate the component, the sensor system comprising: a sensor; a plurality of divergent light sources in the sensor arranged to illuminate a component area in the sensor; a detector disposed relative to the light sources such that when the component is at least partially disposed in the component region, the component shields at least some of the illumination of at least one of the plurality of divergent light sources to create a shadow of at least a portion of the component forming on the detector, the detector being suitable while the component is rotating to provide multiple detector outputs; an optic inserted between a device region and the plurality of divergent light sources to reduce the divergence of the light ...

Surface-bound antigen binding portions of antibodies that bind to ctla-4 and cd28 and uses therefor

System and methods for providing compatibility across multiple versions of a software system

A system and methods that provide compatibility across multiple versions of a software system, such as an execution engine or run-time system, are disclose. The software system is structured so that it includes a master module, and one or more helper modules, each helper module being capable of processing applets (i.e., programs or content) that require a particular version of the software system. Each time an applet is to be processed by the software system, the master module determines which helper module should be used to process the applet, and starts the selected helper module if necessary. Compatibility with additional versions can be provided by providing additional helper modules. Because numerous helper modules may be executed simultaneously, applets that require different versions of the software system may be processed simultaneously.

Human single-chain t cell receptors

A soluble human single-chain T cell receptor (TCR) having the structure: Vα2-L-Vβ or Vβ-L-Vα2, wherein L is a linker peptide that links Vβ with Vα, Vβ is a TCR variable β region, and Vα2 is a TCR variable α region of the family 2 is provided. The provided scTCR is useful for many purposes, including the treatment of cancer, viral diseases and autoimmune diseases.

Allogeneic histocompatibility complexes as mediators of cell destruction

Methods are disclosed of producing and using Major Histocompatibility Complex (MHC) proteins, in native, truncated of fusion forms, complexed with MHC binding peptides and conjugated to targeting molecules, in order to target selected cells in a subject for T cell mediated destruction in vivo. The MHC proteins are chosen so to be allogeneic with respect to the host and the targeting molecules are chosen to selectively bind to the target cells. Pharmaceutical preparations including the targeted peptide-MHC complexes are also disclosed.

Yeast cell surface display of proteins and uses thereof

The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. Attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo . T cell receptor mutants that are efficiently displayed on the yeast cell surface can be isolated by this method, providing a means of altering T cell receptor binding affinity and specificity by library screening. The selection method also identifies proteins with enhanced phenotypic characteristics, proteins that are displayed at higher levels, proteins that are secreted at higher efficiency and proteins that are more stable.

Engineered high-affinity human t cell receptors

T cell receptors (TCRs) that have higher affinity for the Survivin antigen are provided. The high affinity TCRs were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the engineered TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting.

Hochaffine t-zellrezeptorproteine und verfahren

Cancer treatment with 237 car-t cell based therapeutics recognizing the tn epitope

The disclosure provides Tn epitope- specific chimeric antigen receptors and scFvs, including soluble scFvs and multimeric scFvs, as well as methods of identifying cancer subjects and cancer subject sub-populations amenable to anti-Tn immunotherapy and methods of treating cancer.

Receptores de célula t humana de alta afinidade geneticamente modificados e seus usos

RECEPTORES DE CÉLULA T HUMANA DE ALTA AFINIDADE GENETICAMENTE MODIFICADOS. Trata-se de receptores de célula T (TCRs) que têm afinidade maior para o antígeno de survivina. Os TCRs de alta afinidade foram modificados geneticamente através da geração de bibliotecas de mutação de TCRs em um formato de cadeia única, seguida da seleção por afinidade e estabilidade aprimorada, sobre a superfície de levedura (isto é, evolução dirigida). Nas modalidades, os TCRs geneticamente modificados podem ser usados na forma solúvel para a entrega alvejada in vivo, ou como genes introduzidos em células T em uma configuração de célula T adotiva.

Verwendung von verdrängtem syngas zur regeneration von scr-katalysator

System and method supporting property values as options

To support values of properties, a class includes fields to support values in preallocated memory space and with an option data structure which supports, in instances of the class, references to option values without preallocation of memory space. The field and option values are accessed in an instance object of the class using expressions of the same syntactic form.

System und verfahren zur unterstützung von optionentypenkontrolle

Integrated content language for use on the web

The present invention provides apparatus and methods for developing and delivering interactive Web pages and Web-based applications. More particularly, the invention provides content language (10), in which intermixed with programmed content (12) (i.e., code) that defines program behavior. The content and programmed content are expressed in a single content language, using a single linguistic framework having consistent syntax and semantics. This content language includes a number of features, including an ability to handle content, including programmed content as data. In a preferred embodiment, the content language uses an programming constructs, and an infix syntax (15) for expressions. The content language of the present invention may be used to develop interactive web content that may be executed and displayed within a Web browser that includes a plug-in containing a language processor (i.e., a compiler or interpreter) for the content language. A stand-alone language processor for the content language may also be used to execute and display content written in the content language.

System and method supporting plural option data structures

To support values of properties, a class includes fields to support values in preallocated memory space and with an option data structure which supports, in instances of the class, references to option values without preallocation of memory space. The field and option values are accessed in an instance object of the class using expressions of the same syntactic form. During compliation, the compiler checks the type of an option value against a type description within the option data structure. If a value has not been set for an instance object, a get operation results in getting of the default value for the class. Different classes may support different forms of data structures such as a linked list or a hash table. During compilation, a method call to an object is encoded without regard to the form of the option data structure. When an option value is changed, a change handler identified by an option binding of the data structure is processed. That option binding may be located by first searching a mapping data structure for a previously computed mapping to the option binding or by computing the mapping to the option binding. An option value may be set in an option data structure from an initialization expression which includes the name of the option value and, as an argument, the option vale. Nonlocal option values may be applied to plural objects in a nonlocal option hierarchy such as a graphical hierarchy.

Persistent data storage for client computer software programs

Persistent data storage for client computer software programs is provided using a repository that enables client computer software programs to store data securely on a client computer system, subsequently retrieve that data, and optionally share the data in a controlled fashion with authorized client computer software programs. The present invention can be used by both trusted and untrusted client computer software programs that either reside locally on the client computer system or are downloaded from a server computer system. A benefit of the present invention is that it allows untrusted applets to have access to persistent storage without compromising the integrity of the client computer system. Since the present invention controls persistent storage space, not the programmer of the client computer software program, the burden of insuring storage integrity and security is removed from the programmer. In addition to basic integrity and security features, the present invention provides automatic data format conversion for a client computer software programs reading/writing data in the data repository, a commit operation to force in-memory repository data to disk, and automatic expiration to delete the repository data to disk, and automatic expiration to delete the repository after a predetermined time period. The repository can be pre-limited to a maximum size and shared among multiple client computer software programs.

System and methods for providing compatibility across multiple versions of a software system

System and method supporting nonlocal values

To support values of properties, a class includes fields to support values in preallocated memory space and with an option data structure which supports, in instances of the class, references to option values without preallocation of memory space. The field and option values are accessed in an instance object of the class using expressions of the same syntactic form. During compilation, the compiler checks the type of an option value against a type description within the option data structure. If a value has not been set for an instance object, a get operation results in getting of the default value for the class. Different classes may support different forms of data structures such as a linked list or a hash table. During compilation, a method call to an object is encoded without regard to the form of the option data structure. When an option value is changed, a change handler identified by an option binding of the data structure is processed. That option binding may be located by first searching a mapping data structure for a previously computed mapping to the option binding or by computing the mapping to the option binding. An option value may be set in an option data structure from an initialization expression which includes the name of the option value and, as an argument, the option value. Nonlocal option values may be applied to plural objects in a nonlocal option hierarchy such as a graphical hierarchy.

System and method supporting plural option data structures

Persistent data storage for client computer software programs

Hybrid privilege enforcement in a restricted execution environment

A system and method for static and dynamic enforcement of access to resources through a runtime engine allows optimal selection of the enforcement method to improve performance, security, and maintainability of an execution environment. A trust state indicative of permitted resources is defined. Access to resources is provided by invoking particular function instantiations in the runtime engine. Binding of an executable entity to instantiations in the runtime engine occurs selectively during static enforcement based on the trust state. Runtime checks of the trust state by the executable entity occurs during dynamic enforcement. If the trust state does not correspond to a desired resource, access to that resource is prevented.

Hybrid privilege enforcement in a restricted execution environment

A system and method for static and dynamic enforcement of access to resources through a runtime engine allows optimal selection of the enforcement method to improve performance, security, and maintainability of an execution environment. A trust state indicative of permitted resources is defined. Access to resources is provided by invoking particular function instantiations in the runtime engine. Binding of an executable entity to instantiations in the runtime engine occurs selectively during static enforcement based on the trust state. Runtime checks of the trust state by the executable entity occurs during dynamic enforcement. If the trust state does not correspond to a desired resource, access to that resource is prevented.