Procedure for the production of a sturdy and of hypotensiven materials free plasma protein

The invention refers to a procedure for the production of a sturdy and of hypotemiv working materials free plasma protein. in particular such a sturdy plasma protein, which has and for infusions which can be accomplished rapidly be used can no depressive effect. Lately in the context of medical treatment plasma protein on the infusion way was given increasingly, instead of making blood transfusions. Hiebei could be obtained considerable therapeutic effects, if with the occurrence of a shock, by Hypoproteinämie and with excessive Blutun - towards during operations a sturdy plasma protein free of active hepatitis virus were intravenously given. Such a plasma protein was however not completely free from hypotensiv working materials. It is now a goal of the invention of creating a procedure for the production of a sturdy plasma protein and of manufacturing hiebei from each depressive effect free and for infusions which can be accomplished rapidly useful plasma protein. This succeeds to be essentially removed, if in accordance with the invention from blood pressure-lowering effect determines a possessing and, elektrophoretisch, from an albumin parliamentary group for existing sturdy plasma protein, which had been won from human blood or Plazenta and heated up hiebei during the production steps or afterwards on a temperature from 57 to 60°C, a molecular weight from 1000 to 10,000 possessing Peptide by Ultrafiltrieren or gel filtration or after an adsorption process. The fractionation of mixtures of high-molecular materials after their Molekularge icht by means of the gel filtration is based on that in the pores of a gel, for example an transverseinterlaced Dextran, solvent-poured by water or another polar, only materials with underneath an upper limit value of lying molecular weight determined by the Porengröße of the gel are held back and from there with filtering a solution of a material with underneath the mentioned upper limit value and materials with a molecular weight of contained material mixture lying above the mentioned upper limit value from the gel layer only the materials with a molecular weight containing solution lying above the mentioned upper limit value withdraw and afterwards from the Ge1 successively further materials according to falling molecular weight regulatorily to be washed to be able. The Ultrafiltrieren is similar in principle to the gel filtering, however hiebei a porous diaphragm consisting of a gel is used, which lets materials through with a molecular weight lying underneath an upper limit value determined by the Porengröße, so that from the print page of the diaphragm the materials with a molecular weight lying above the mentioned upper limit value containing solution can be taken off and from the other side of the diaphragm the materials with a molecular weight containing solution lying underneath the mentioned upper limit value. Invention in accordance with [...] producible sturdy plasma protein does not exist the main thing after made of albumin and to the smaller part made of A-Globulin and 8-GlobUlin.Die heat resistant A-Globuline and 8-Globuline is distinguishable from the albumin by Ultrazentrifugieren analytically. For this reason the stable plasma $5 protein is not identified when examining according to elektrophoretischen methods as pure albumin, however it is characterized by only one vertex, which is alike to the albumin with the Ultrazentrifugieren. Details result from the designs. Fig. la a blood pressure curve shows, which in the case of intravenous administration of 188 mg/kg a sturdy plasma protein arises, which from venous blood before heating up on 60°G during 10 h is. Blood pressure lowering: 0%0. Fig. lb a blood pressure curve shows, which in the case of intravenous administration of 188 mg/kg a sturdy plasma protein arises, which was made of venous blood by heating auf60°C up during 10 h. Blutdmcksenkung: ! 1, 5%. Fig. lc a blood pressure curve shows, which in the case of intravenous administration of a sturdy plasma protein arises, that from venous blood by treatment in with Silicagel filled a column after heating up on 60°C during! 0 h received is. Bhtdrucksenkung 0%. Fig. ld with administration of 188 mg/kg human serum albumin resulting blood pressure curve shows. Blood pressure lowering: 0%. Fig. le still serves a freon for the comparison of the contractions of the smooth musculature after administration of 10 ng Bradykinin (A), after administration of a solution out venous blood of a won sturdy plasma protein (B) and administration of a 5%oigenLösung out venous blood of a won and sturdy plasma protein (C) treated with Silicagel. Fig. 2a shows a blood pressure curve, which in the case of intravenous administration of 188 mg/kg a plasma protein arises, which had been received to heating auf60°C up directly from Plazenta or during 1 h in presence from butter acid. Blood pressure lowering: 0%0. Fig. 2b shows a blood pressure curve, which in the case of intravenous administration of 188 mg/kg a sturdy plasma protein results, which from human Plazenta was won. Blood pressure lowering: 19! %. Fig. 2C shows a blood pressure curve, which in the case of administration of 188 mg/kg a sturdy plasma protein results, which from human Plazenta by BehandeIn with Silicagel had been received. Blood pressure lowering: 0 o. Fig. 2d serves a 5% RST for the comparison of the contractions of the smooth musculature after Vera breichung - sung a out human Plazenta by treatment with Silicagel of won sturdy plasma protein (D), after administration of the same however not treated solution (e), after administration 8% igen solution memchllchen serum albumin (f) and after Verabreiehung of Bradykinin in a quantity of 20 ng (g), of g 10 Hg (h) und5 Hg (i). In the above figures by the symbols “A” and “B” were suggested that the administration was made between these times “A” and “B”. Fig. 3 illustrates those parliamentary groups, which with the gel filtering of the stable Plasmaproteim received in human Plazenta over Sephadex G-g0 (trade name of the company Pharmacia, Upsala, Sweden, for! 0 hydrophilic and insoluble molecular sieve from quervemetztem Dextran, which is used for chromatographische purposes) to arise. With the gel filtering the following conditions of work were selected: Dimensions of the column: cm diameter cm height of volumes of the column filling: given up password: 11 solution of the sturdy plasma protein quantity of the given up solution: 70 ml volumes of each sample tube: i0 ml Eluiermittel: 0, 5 m-NaCl Eluiergeschwindigkeit: 4 valley/min. Already different procedures for the production of a heat resistant plasma protein from fresh memchlichem blood were practically accomplished, however it prepares to receive numerous difficulties large quantities of fresh blood plasma of healthy persons. Since now Plazenta is nevertheless rich at groove and it is easily processable is usually rejected, efforts were already undertaken to win plasma protein from Plazenta however appropriate procedures were not practically accomplished yet. For the production of a sturdy plasma protein from plasma already different procedures were suggested. (1) one this-proceed-places modify-proceed vonCohn the Indian Japanese patent publication No. 5297/60 method indicated in sixth place to the Fraktioniesen of a plasma protein. With - its procedure is precipitated less from blood plasma by means of 25 to 28%igem ethanol at a pH value of 4, 3 to 4, 7, at a temperature from -2 to -20°C and with an ion strength of 1, 9 or a mixture from albumin, A-Globulin and 3-Globulin, on which that released kind of such from the entire Fibrinogen and from entire 7 " Gl°bulin sturdy plasma protein is separated and by 10stündiges heating up on a temperature of 60°C if necessary in this plasma protein available hepatitis virus inactivated. In such a way manufactured sturdy plasma protein possesses however by Fig. Ib illustrated blood pressure-lowering effect; in contrast hiezu however the plasma protein before heating up did not possess this blood pressure-lowering effect (see Fig. la). (2) gladly [...] the Japanese patent publication No. 16 041/68 a sturdy plasma protein from large mixing a fabric excerpt containing at hemoglobin or of large quantities of hemoglobin containing blood was already made, as butter acid were eye-set and by heating a pH value of 4, 5 to 5, 5 large-mixed possessing the mixture up on 57 to 60 °C instable Globine separated at Hamoglobin containing excerpt. In detail with this procedure for the production from albumin, A - Globulin and - Globulin existing Plasmaproteim from the supernatant liquid a y-Globulinenthaltende parliamentary group from that one large-mixes is stopped finally at hemoglobin containing plasma or Plazentaextrakt precipitated, by the supernatant liquid Butters äure up to reaching a concentration from 2 to 6% one adds, afterwards the pH value to 4, 5 to 5, and the solution on 57 to 60°C is heated up, around the w ärmelabilen Globine auszufäUen. The sturdy plasma protein manufactured in this procedure possesses, as by Fig. 2b is described, blut00 printinglowering effect, against what the solution available before heating up on 58 to 60°C did not possess blutdrueksenkende effect (see Fig. 2a). Bland and coworkers observed blutdrucksenkendeWtrkung in accordance with the above procedure (!) manufactured stable Plasmaproteim, as it with surgical interferences at hearts at least! 00 ml a 5% solution of the sturdy plasma protein injected, and zw. sees this effect 4 min pointed to the injection (J.H.L. Bland, N.B. Laver and E. Lowenstein, Hypotension Due ton of Five Percent plasma protein Fractions, new England. L Med. 286 S.109). Also Harrison and coworkers reported on the blood pressure-lowering effect in the above procedure (1) manufactured Plasmaproteim with administration of the same on the Infusiomwege (G.A. Harrison, M.Robinson, R.V. Stacey, C.H. McCulloch, T.A. Torda and J.S. WRIGHT, Hypotemive Effects OF Stable plasma protein Solution. A preliminary Communication, Medical J. Australia 1040 Bd. 2; G.A. Harrison, T.A. Torda, and P. ship, Hypotensive Effects OF Stable plasma protein Solution. A preliminary Communication, MedicalJ. Australia 1308, Bd.). At present can in the procedure mentioned above (1) manufactured sturdy plasma protein clinically only into the Vene to be instilled, since with rapid infusion of this protein the danger exists to strong blood pressure lowering. During the rapid advancement of the surgical methods however after a need on the infusion way of rapidly givable plasma protein ever more one strengthens. The patent owner looked for now for ways from solutions one for the above procedures (1) and (2) manufactured stable Plasmapmteins the hypotensiv working substance to separate and stated hiebei that it concerns with this hypotensiv working substance a Peptid possessing a molecular weight from 1000 to 10,000, which with everyone of the procedures indicated above (I) and (2) during heating up on Tempe - raturen from 56 to 60 °C develops. This hypotensiv working Peptid effectuation also the contraction of the smooth musculature, is inactivated however by Carboxypeptidase B. It was now found that this hypoten - siv working Peptid in front sturdy plasma protein can be separated by gel filtration or U1trafiltration, if was ensured that the molecular weight of the hypotemiv working Peptids is smaller than that one of the sturdy plasma protein. Beyond that also found that this hypotensiv working Peptid is adsorbable at cation exchangers and inorganic Adsorbentien also, however the sturdy plasma protein at such Adsorbentien is not adsorbed, so that the hypotensiv working Peptid can be separated also in this way from the sturdy plasma protein. It was further found that as soon as from in the upper mentioned procedure (1) by 10stündiges heating up on 60°C manufactured sturdy plasma protein the hypotensiv working Peptid into according to invention-wise, developed during heating up as by-product, separated, this hypotensiv working Peptid is with later heating of the stable Plasmaproteim up on temperatures around 60 °C any longer does not develop. Beyond that the hypotensiv working Peptid does not develop also any longer if in the procedure mentioned above (2) manufactured sturdy plasma protein the if necessary existing hepatitis virus by 10stündiges heating up on 60°C one inactivates, if this was separated before hypotenstv working Peptid in way according to invention (see Fig. 2C). The procedure according to invention is based thus on the mentioned new realizations and makes possible it from blood pressure-lowering effect free and for rapid infusion suitable plasma protein from human blood and/or of Plazentaextrakt to make. After the procedure according to invention it is thus possible, 10,000 possessing sturdy plasma protein freely from each blood pressure-lowering effect by separating for an in molecular weight of 1000 to win hypotemiv affecting Peptids from a protein which after electrical - phoretischen methods determines, essentially from albumin consists and of human blood or human fabric was herausfraktioniett and during the Frakttonierem or 57 to ü0°C heated up afterwards. Since producible the according to invention sturdy plasma protein does not contain a hypotensiv working Peptid, also then no danger of an excessive Blutdmcksenkung exists with patents, which this sturdy plasma protein is instilled not into the Venen, but given on the infusion way rapidly. The clinical Auwendbarkeit of sturdy plasma protein is considerably increased thereby. In accordance with the invention not only pure plasma, but also large quantities of hemoglobin containing plasma and/or Plazentaextrakt can be used for the production of the sturdy plasma protein, with which sturdy plasma protein substantially landlord - becomes more schaftlicher producible. In the case of production according to invention of the plasma protein from blood, needed for the procedure, the sixth method already mentioned can vonCohn for out fractionation of plasma protein [a procedure, by which with fractionation by means of ethanol at low temperatures zinc one introduces (see Trade Union of German Employees-read. M. Sargener et al. : Vox. Sunginis, Bd. 5 P. 272) or a procedure, with which the ion strength is reduced by plasma by means of an ion exchange resin, around instable Globuline auszufäIlen (see Hans Nichman et al. Vox.Sunginis, Bd. 3 P. 184) to be used. Plasma protein can be placed made of plasma or large mixing a Plazentaextrakt containing at hemoglobin also in the procedure in accordance with the Japanese patent publication No. 16 041/68 indicated above ago -. In the special plasma protein can be made of Plazenta in the following way. At low temperature frozen Plazenta is roughly cut up in a EismühIe and continued to divide in a meat mill, on which 100 thread - parts of in such a way cut up Plazenta with 200 ml one percent saline solution to be shifted and after 30 min during extracting the Plazentaextrakt representing liquid is abzentrifugiert. Then Ammonsulfat is added ge to the received Plazentaextrakt in such Men - that a Ammonsulfatkonzentration from 35 to 40% results. If the possibility exists to hold for the Plazentaextrakt on sufficiently low temperature to this Plazentaextrakt also ethanol in a quantity of 25 thread - parts per 100 thread - can parts of Plazentaextrakt be added and hiebei the developing precipitation be separated. The larger part of hemoglobin remains in hiebei the received clear solution, which can be subjected directly to the next process step, and zw. a treatment with organic acid, however appropriately with Ammomulfatversetzt becomes, because the concentration of the protein in the solution is low to precipitate a raw albumin parliamentary group. The received precipitation is filtered off and then in 3 to 4 1 water per kg precipitation solved. In in such a way received raw albumin parliamentary group if necessary existing Ammonsulfat does not disturb with the treatment with an organic acid, which can be made later. From in such a way received solution by adding an organic acid the hemoglobin still solved in this solution is precipitated, which is filtered off. Hemoglobin is precipitated the more to a large extent, the added more organic acid of the solution, however, although with Grölerei quantity of added organic acid separating the desired protein is facilitated, with additive of too much organic acid the yield at sturdy plasma protein is reduced thereby that during the later thermal treatment also albumin is precipitated. For this reason it is indicated, to add which with the organic acid solution so much organic acid which can be shifted that the finally received solution contains 2 to 6% at organic acid. In order to precipitate the Hämoglobfn completely, is to the pH " value of the solution special attention to be given. The quantity of hemoglobin will with reducing the pH value of the solution of the neutral value to 5, loosened in the solution, 5 considerably reduces, whereby with lowering the pH value on 4,3 hemoglobin is practically completely precipitated. It is however indicated to the solution, the pH value to a value between 4, 5 and 5 to bring 5 since with lowering the pH value on still low-worth also those-exploit at albumin drops. The received solution is then gradually warmed up, whereby at temperatures around 50°C and over it warm-inconsistant Globuline begin to fail. The temperature of the solution is held sodarm certain time to a value between fit and for 60°C, whereby bei60°C instable Globulin is usually precipitated within 1 to 2 min, however is more favorable it to hold the selected temperature during 15 to 30 min in order to make sure that all Globulin is precipitated; this is easily possible, since the albumin is heat resistant, thus also the sturdy plasma protein. To precipitate subsequently, to heating up the received solution cooled down on ambient temperature, on which the available precipitation cool after the off were filtered off and to the received filtrate, which contains no more hemoglobin, per litre 520 g Ammonsulfat added, in order the proteins completely. The received precipitation is filtered off and dialysiert in a cellophane pipe against cold water, in order to separate from the protein Ammomulfat and organic acid. It becomes in such a way 5, 0 to 6, 5 thread - °70 sturdy plasma protein containing solution receive, from which the hypotensiv working substance can be separated in the way indicated above by Hindurchleiten the solution by with more lonenaustanscher or an inorganic adsorbent filled column or by gel filtration or by Ultrafiltrieren, in order to arrive at one of hypotensiv working substance to released sturdy plasma protein. In the context of the procedure according to invention venqendbare generally used cation exchanger is Ionenaustan such as Carboxymethylcellulose, Carboxymethylsephadex (trade name of the company Phamaacia Fine chemicals for a hydrophilic and insoluble KationenaustauscherausquervemetztemDextran) and amberlites CG-50 (trade name for a weakly sour cation exchange resin). As inorganic Adsorbentien Silicagel or alumina gel can be used in the context of the procedure according to invention with advantage. At the lonenaustauschharzen and/or inorganic Adsorbentien mentioned above the hypotensiv working substance contained in the sturdy plasma protein is absorbed preferably thereby that on ion exchange resin brought with 0, 2507oiger sodium chlorid solution in the equilibrium and/or inorganic adsorbent filled column the too reirägende solution of the plasma protein with a speed of 50 to 150 ml/h. cm z is indicated, whereby preferably at temperatures from 4 to 6°C one works. If in the context of the procedure according to invention the gel filtration is used, hiefür well-known methods can be used. Useful methods are among other things indicated in the following literature places. 1. Determination OF molecular weights OF of protein by gel filtration on Sephadex. Anal. Chem. 35 P. 1950 to 1958, Whitaker, J.R. 2. Estimation OF the molecular weights OF of protein by Sephadex gel flltration. Biochem. J. 91 P. 222 to 233, Andrew, P. 3. A theory OF gel filtration and its experimental verification, J. Chromatog. 14 P. 317 to 330, Laurent. T.C., Killander, J. 4, Estimation OF molecular size OF of peptide by gel filtration, Biochem. J. 95 9 P, Carnegie P is ultraflltriert. IL of case in the context of the procedure according to invention, can be worked under the conditions indicated in the following literature places. 1. Separation and purification OF biological material by ultrafiltration, H.J. Bixler, P. W. house flax, L.M. NelsenMay., Nat. Meeting. To. Inst. OF chem. one close. Smart country. 2. Rem oval OF lgM would freeze serum by ultrafiltration, C.J. Van Ost. P.M. Bronson, Anal. Biochem., 36, S. 464. 3. Ultrafiltration OF humanly serum, evidence OF low molecular weight cholinesterase activity (in French), D. Boutin J. Bmdeur, Rer. CAN. Biol. 29 (2), 187 [June 1970]. Can be added to the sturdy plasma protein released in this way from hypotensiv working substance stabilizers, beispielsweiseN Acetyltryptophan and Natriumcaprylat, in such quantities that the concentration of the stabilizers in the solution amounts to 0, 0032 to 0, 0046 m, whereby 10, 5 to 14, 5 h to 59 to 61°C can be heated up. lm available case the blutdmcksenkende effect was always determined to a body weight male dogs of a crossing different dog races fully developed by approximately 8 kg of possessing, put to how that dog according to the Anästhesieren with urethane on the backs and into the left carotid artery of the dog eineKanüleeingefllhrtwurde, in order to be able to tune by means of a multiple writer the blood pressure in the carotid artery. The connection which can be examined became the Hünd over to the right Hüftvene befe - stigten Katheder supplied. The Blutdmcksenkung was determined from the following equation, whereby old “average blood pressure was inserted after administration of the connection” the lowest blood pressure measured after administration of the connection into the equation and the “average blood pressure was used before administration of the connection” as base factor. Blood pressure lowering in % Dumhschnittlicher average blood pressure before blood pressure after administration abreichung the VerVerbindun connection average blood pressure before administration of the connection × 100 the effect on the smooth musculature was determined at the Uterus by rats according to the method by Magnus, whereby the Uterus was used by virgin female rats. A body weight of approximately 150 g possessing female rats of the slot Wister 18 h before removing the Uterus 5 mg Diöstradiol were given, on which 18 h were beheaded later the rats and before removing the Uterus the blood flow out were let. As nutritive solution with air satisfied and 0, 1 mg per 100 ml atropine sulfate enthalende solution after Jalon were used. The volume of the nutritive solution in the organ amounted to 8, 6 mI, the volume of the sample solution amounted to 0, 4 ml and the Re.aktiomzeit amounted to 90 seconds. The procedure according to invention is more near described in the following on the basis of remark examples. Example I: After the patent specification No. Japanese of Cohninder 5297/60 fractionating method indicated in sixth place was won from I00 1 memchlichem plasma a sturdy plasma protein. For this purpose the plasma was brought on a pH value of 7, 2, then with 53, to 3%oigem cold Äth - anol bit for reaching a concentration of 8% transferred and schliefllich with -2°C centrifuges, in order to separate the Fibrinogen. Further cold ethanol up to reaching a concentration of 21% was abzentrifugiert added to the received clear solution, on which the pH value of the received solution auf6, 8 adjusted, then the temperature of the solution gradually to -6 °C lowered and schliefllich), - Globulin. In such a way received clear solution was shifted I at a pH value of 4, 7 and with lonemtärke of 0, at a temperature of -6°C with as much 25%igem ethanol that a mixture from albumin, A was precipitated - G1obulin and B-Globulin. The precipitated precipitation was separated from the solution and afterwards more gefriergetrockner, with which 3, 2 kg of a dry and powdery Proteim became to receive. The received powder was converted with distilled water to a solution 5% at protein, which N-Acetyltryptophan and Natriumcaprylat were then added as stabilizers in such quantity that a concentration of 0, 04 m resulted. In such a way received solution was then warmed up I0 h to 60°C and given up anschlieflend on a column filled with Sillcagel, whereby from the column a plasma protein flowed off a containing parliamentary group. The hypotemiv working substance was adsorbed hiebei at the Silicagel. The parliamentary group flowing out from the column contained about 95% of the ursprtinglich available proteins in form of a sturdy plasma protein. The received experimental results are in the following table I and in the Fig. la to le indicated. Table I solution of the sturdy plasma protein before the treatment with Silicagel in a column solution of the sturdy plasma protein after the treatment with Silieagel in a column yield at sturdy plasma protein before and/or after the treatment with Sillcagel, blood pressure-lowering effect in dogs and contracting effect on the smooth musculature of the Uterus of rats rehearse OD 30, 8 31.2 80, 0 30, yield at protein (lOO%) (the contracting effect is located to 1oo%) 95 o 93, 5% blood pressure-lowering effect with dog EN 4Kontrahierende effect on the smooth musculature of the Uterus of rats in the above table I and into the following table [...] and 1II the column concerned the indication in “- -” for one by 25 to 125 mg Bradykinin per ml contained solution erzielbamnKontraktion gIetche a contraction and the indication “-” for a that-been missing contraction, against what in the column concerned the blutdmcksenkende effect the indication is located ““for a waste of the arterial Blutdmckes around 11, 5 to 25, 0% and the indication”--. “being missing a blood pressure decrease means. According to a elektrophoretisch accomplished analysis contains the sturdy plasma protein 88, 5%o A1bumin, 7, 5% A-GIobulin and 4, 0%/3 - Globulin. 5% solutions of the sturdy plasma protein can rapidly on the infusion way given werdeu, without having any blood pressure-lowering effect. Example 2: , The sturdy plasma protein powdery manufactured according to the sixth modified method of Cohnausmenschlichem plasma in accordance with example 1 was converted with distilled water to 5% protein a containing solution, N-Acetyltryptophan and Natriumcaprylat in such quantity were more softly then added that a final concentration of 0, 04 m resulted. Subsequently, the solution 10h on 60°C was heated up. After heating up the solution of the PIasmaproteins became using a UM10-Membran (manufactured Amicon CO. The USA) ultrafiltriert, whereby the hypotensivwirkende Peptid was filtered off by the Plasmaprotcin. The received results are indicated in table [...]. Solution of the stable Plasrcaproteins before the Ultrafiltriereu solution of the sturdy plasma protein after the Ultrafiltrteren table 11 blood pressure-lowering effect and kontrahiemnde effect on the smooth musculature of a solution of the sturdy plasma protein before and after the Ultrafiltrteten sample 3 4 3 4 yield at protein 100% 100% blood pressure-lowering effect in dogs H H contracting effect on the smooth musculature of rats the protein concentration of the solution of the Plasrcaproteins released by Ultrafiltrieren from the hypotensiv working Peptid was stopped to 5%, on which the solution with N-Acetylttyptophan and Natfiurccaprylat up to reaching a Endkonzentrationvon0,0Bm was shifted. Hiebei a stable Plasrcaprotein in form of a solution, rapidly first-richable on the infusion way, was received, which did not contain a hypotensiv working substance. Example 3: To finely keep immediately after the Anliefetung frozen and in this condition stored Plazenta husbands, on which to 20 kg of verrcahlener Plazenta 40 were given to 1 of a one percent saline solution, in order blood components a containing excerpt. From this excerpt one was precipitated - Globulin containing parliamentary group, on which from 2 to 6% it was added to the received clear liquid butter acid up to reaching a final concentration and then the pH value of the solution treated with acid was stopped to 4, 5 to 5, 5. Subsequently, the solution still 1 to 2 h to 57 to 60°C was warmed up and the developed precipitation by centrifugation was separated. Arcmortsulfat up to reaching a concentration of 75% was added to the received clear solution, with which the entire proteins were precipitated. So the plasma protein received in form of a paste in a Dialysierrohr was brought in and in it with 4°C 24 h against flowing water dialysiert (this function corresponds to the procedure to here gercäß the Japanese patent specification No. 16041/68). In such a way manufactured solution of the Plasrcaproteins was brought on a pH value of 7, 0 and then brought on a column filled with Carboxyrcethylsephadex given up, before with 0, 05 rc-Phosphatpuffer (pH = 6, 8 to 7, 0), which at Natriurcchlorid 0, 1 rcolar was, in the equilibrium was. Subsequently, the column with the buffer mentioned was eluiert. In the Eluat about 90% were contained of the Plasrcaproteins given up on the column, however the hypotensiv working Peptide at the column filling had been adsorbed. The received results become by the following table llI and by the Fig. 2a to 2d describes. Table [...] stable Hasmaprotein before treating with Carboxymethylsephadex in a column sturdy plasma protein according to treating with Carboxymethylsephadex in a Kolorme yield at sturdy plasma protein, blood pressure-lowering effect in dogs and contracting effect on the smooth musculature of the Uterus of rats forwards and according to dern regenerating the solution in the column sample No. filled with Carboxymethylsephadex. ODz80 32, 8 36, 7 32, 0 35, 2 yield at protein (IO o) (lOO%) 91, 0% 89, 5% blood pressure-lowering effect in dogs contracting effect on the smooth musculature of the Uterus of rats the solution of the plasma protein received by treatment with Carboxymethylsephadex adjusted to a protein concentration of 5%, on which the received solution with N-Acetyltryptophan and Natriumcaprylat was shifted, to itself a final concentration of 0, 04 m resulted in. The now available solution was heated up still 10 h on 60°C, with which on that lnfusiomwege a rapidly givable and of hypotensiv working Peptid completely free sturdy plasma protein became to receive. The received sturdy plasma protein consisted to 95% of albumin, A-Globulin and t3-Globulin and was present as 5% ProteinIösung. The yield amounted to about 2, 2 g per litre of Plazenta. 13 e i s p i e 1 4: After the function indicated in example 3 out fatter and immediately angelie became - after delivery of frozen Plazenta in the fresh condition, and zw. by addition of the liquid and the gel filtering over Sephadex G-50 a plasma protein liberating from hypotemiv working substance (low molecular weight) (high molecular weight), centrifuged by Buttersäute, heating up on 57 to 60°C, centrifugation of the liquid, Dialysieren, manufactured (see Fig. 3). The protein concentration of the received solution of the sturdy plasma protein was stopped to 5%, on which N-Acetylttyptophan and Natrfumcaprylat up to reaching a final concentration were added to the received solution by 0, 04 m. The received solution was then heated up still 10 h on 60°C, received with what on that lnfusiomwege rapidly givable and no hypotemiv working Peptid containing sturdy plasma protein.