CS261849B1 - A method of isolating plasma proteins from ram plasma or serum - Google Patents

A method of isolating plasma proteins from ram plasma or serum Download PDF

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CS261849B1
CS261849B1 CS876284A CS628487A CS261849B1 CS 261849 B1 CS261849 B1 CS 261849B1 CS 876284 A CS876284 A CS 876284A CS 628487 A CS628487 A CS 628487A CS 261849 B1 CS261849 B1 CS 261849B1
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Czechoslovakia
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plasma
serum
plasma proteins
isolating
proteins
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CS876284A
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Czech (cs)
Slovak (sk)
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CS628487A1 (en
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Tamara Mvdr Kohlmayerova
Ivan Doc Mvdr Csc Mikula
Juraj Mvdr Csc Balascak
Tomas Mvdr Dravecky
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Tamara Mvdr Kohlmayerova
Mikula Ivan
Juraj Mvdr Csc Balascak
Tomas Mvdr Dravecky
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Priority to CS876284A priority Critical patent/CS261849B1/en
Publication of CS628487A1 publication Critical patent/CS628487A1/en
Publication of CS261849B1 publication Critical patent/CS261849B1/en

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Abstract

Riešenie možno využit v imuno-bltologickej výrobě na izoláciu plazmatických bielIkovín z baranej plazmy alebo séra. Účelom je příprava výrobku o širšom spektre stabilizovaných plazmatických bielkovín určeného pre liečenie chorých zvierat. Zvýšenie účinku sa dosahuje tým, že za vhodných podmienok pH, teploty a kyseliny ka- 'prylovej sú izolované plazmatické bielkovihy obsahujúce 40 až 50 % imunoglobulínov a 50 až 60 % sériového albuminu.The solution can be used in immuno-bltological production for the isolation of plasma proteins from sheep plasma or serum. The purpose is to prepare a product with a wider spectrum of stabilized plasma proteins intended for the treatment of sick animals. The increase in effect is achieved by isolating plasma proteins containing 40 to 50% immunoglobulins and 50 to 60% serum albumin under appropriate conditions of pH, temperature and caprylic acid.

Description

Vynález sa týká sposobu izolácie plazmatických bielkovín z baranej plazmy alebo 'séra, pri ktorom sa za vhodných podmienok pH, teploty a koncentrácie kyseliny kaprylovej izolujú plazmatické bielkoviny obsahujúce v prevážnej miere imunoglobulíny a albumin. V súčasnom období sa v priemyselnom měřítku izolujú z hovadzej, konskej a prasečej plazmy, popřípadě séra niektoré plazmatické bielkoviny, ktoré sa používajú pri liečení chorých zvierat. Ide hlavně o izoláciu a přípravu imunoglobulínu (Gamma Globulin, (Graeub), The Veterinary Record, *17, 1974; Imunoglobulín zvíraci homologni, Návod na použitie, o. p. OSOL; Gamaglobulín. hovadzí a pre ošípane, Předpis pre výrobu a použitie přípravku č. VPL 01—126— —84, Bioveta n. p. Nitra) a sérového albuminu (Hovězí sérový albumin, PNBI 302/84, Bioveta n. p., Ivanovice n,a Hané; Plazbipn-B,Předpis pre výrobu, kontrola a použitie přípravku č. VPL 01—131—84, Bioveta n. p., Nitra; Mikula I., Rosocha J., Bulík J.: SpoSob izolácie zvieracieho albuminu, PV 1035—85).BACKGROUND OF THE INVENTION The present invention relates to a method for isolating plasma proteins from ram plasma or serum, wherein plasma proteins containing predominantly immunoglobulins and albumin are isolated under appropriate pH, temperature and caprylic acid conditions. At present, some of the plasma proteins used in the treatment of diseased animals are isolated from bovine, equine and porcine plasma or serum, respectively. These are mainly the isolation and preparation of immunoglobulin (Gamma Globulin, (Graeub), The Veterinary Record, * 17, 1974; Immunoglobulin animal homologous, Instructions for use, op OSOL; Gamaglobulin bovine and for pigs; VPL 01—126——84, Bioveta np Nitra) and serum albumin (Bovine Serum Albumin, PNBI 302/84, Bioveta np, Ivanovice n, and Hané; Plazbipn-B, Production, Control and Use Regulation No. VPL 01 —131—84, Bioveta NP, Nitra, Mikula I., Rosocha J., Bulík J .: Isolation of Animal Albumin, PV 1035—85).

Praktické skúsenosti ukázali, že pri Bečení niektorých chorobných stavov zvierat bolo by vhodnejšie podávat širšie spektrum 'stabilizovaných plazmatických bielkovín vo 'formě injekčnéhO roztoku. V minulosti obdobné případy boli riešené aj v humannej medicíně, kde bol vyvinutý preparát obsahujúci zmes albuminu a globulínov pod ná'zvom Stabilný roztok plazmatických bielkovín (Rybák M., Škvařil F., Andrašina J.: Csl. farmácia 3, 137, 1963; Milár a spol.: Stabilný plazmatický roztok bielkovín — Záve'rečná správa 1965; Jusko B., Bulík J.: Sposob přípravy plazmatického roztoku ludských bielkovín, AO 182 921; Kyseliev A. E., From A. A.„ Nikitenko A. A., Rusanov V. M.: Problemi gematologii i perelivania krovi 4, '9, 1967; Bulík J., Banda I., Stachý A.: Sposob izolácie 1'udských plazmatických bielkovín obsahujúcich albumin, AO č. 248 241). i Podstata izolácie plazmatických bielkovín obsahujúcich imunoglobulíny a albumin z baranej plazmy alebo séra spočívá v tom, 'že hodnota pH čerstvej plazmy alebo séra Sa upraví v rozmedzí 4,45 až 4,85 a pri teplote —16 až —j- 18 D,C sa za stálého, miešania přidává koncentrovaná kyselina kapry'lOvá v množstve 2(3 až 26 rol na 1 000 ml Východzej suroviny. Po dokonalej homogenizácii miešaním sa vyprecipitované balastné bielkoviny oddelujú centrifugáciou a supernatant, ktorý obsahuje v prevážnej miere imunoglobulíny a albumin sa čeří filtráciou, dialyzuje oproti 0,45 % roztoku chloridu sodného po dobu 40 až 50 hodin. Příklad 1 1 000 ml baranej plazmy sa vytemperuje na teplotu 4- 16 °C, hodnota pH sa upraví IM kyselinou octovou na 4,65 až 4,75 a stálého miešania sa po kvapkách přidává 25 'mililitrov kOncantrovanej kyseliny kaprylovej. Po dokonalom miešaní v priebehu 3 hodin vyprecipitované balastné bielkoviny sa oddelujú na kyvetovej odstredivke. Sufpennatant sa filtruje cez čeriace vložky a dialyzuje oproti 0„45 % roztoku chloridu Sodného po dobu 4(2 hodin. Získaný roztok Obsahuje 60 % albuminu a 40% imunoglobulínov. 'Příklad 2 2 000 ml baranieho séra sa vytemperuje na teplotu 17 °C, hodnota pH sa upraví 1 M kyselinou octovou na 4,75 až 4,85 a za stálého miešania sa po kvapkách přidává 48 ml koncentrpvanej kyseliny kaprylovej. Po dokonalej homogenizácii miešaním v priebehu 3 hodin sa vyprecipitované balastné bielkoviny oddelujú na kyvetovej odstredivke. Supernatant sa filtruje cez čeriace vložky a dialyzuje oproti 0,45 % roztoku chloridu sodného po dobu 48 hodin. Získaný roztok obsahuje 55 % albuminu a 45 % Imunoglobulínov.Practical experience has shown that, in the wake of some animal disease states, it would be preferable to administer a wider range of stabilized plasma proteins in the form of an injectable solution. In the past, similar cases have also been addressed in human medicine, where a preparation containing a mixture of albumin and globulins was developed under the name Stable Plasma Protein Solution (Rybák M., Škvařil F., Andrašina J .: Csl. Pharmacy 3, 137, 1963; Milár et al .: Stable Plasma Solution of Proteins - Final Report 1965, Jusko B., Bulík J .: Method for Preparation of Plasma Protein Solution, AO 182 921; Kyseliev AE, From AA "Nikitenko AA, Rusanov VM: Problems of Gematology perelivania krovi 4, '9, 1967, Bulík J., Banda I., Stachy A .: The Method of Isolation of 1' Human Plasma Proteins Containing Albumin, AO No. 248 241). The principle of isolation of plasma proteins containing immunoglobulins and albumin from plasma or serum is that the pH of fresh plasma or serum is adjusted between 4.45 and 4.85 and at a temperature of -16 to -18 D, C. concentrated caprylic acid (2 to 26 rolls per 1000 ml of starting material is added with constant agitation). After thorough homogenization by stirring, the precipitated ballast proteins are separated by centrifugation and the supernatant containing predominantly immunoglobulins and albumin is filtered by filtration. dialysing against a 0.45% sodium chloride solution for 40-50 hours Example 1 1000 ml of rammed plasma is brought to 4- 16 ° C, the pH is adjusted to 4.65 to 4.75 with 1 M acetic acid and 25 milliliters of uncanated caprylic acid are added dropwise with constant stirring, and after complete stirring for 3 hours, the precipitated ballast proteins are separated into the Sufpenate is filtered through a pad and dialyzed against a 0 45% sodium chloride solution for 4 (2 hours). Solution obtained Contains 60% albumin and 40% immunoglobulins. EXAMPLE 2 2000 ml of mutant serum are brought to 17 DEG C., the pH is adjusted to 4.75 to 4.85 with 1 M acetic acid and 48 ml of caprylic acid are added dropwise with stirring. After thoroughly homogenizing with stirring for 3 hours, the precipitated ballast proteins are separated on a cuvette centrifuge. The supernatant is filtered through pads and dialyzed against a 0.45% sodium chloride solution for 48 hours. The resulting solution contains 55% albumin and 45% immunoglobulins.

Claims (1)

PREDMETSUBJECT Sposob izolácie plazmatických bielkovín z baranej plazmy alebo séra zražaním kyselinou kaprylovou vyznačujúci sa tým, že pH východzej suroviny sa upraví na hodnotu 4,45 až 4,85 a pri teplete + 16 až + 18 °C sa za stálého miešania přidává koncentrovaná kyselina kaprylová v množstve 23 ažProcess for isolating plasma proteins from plasma or serum by precipitation with caprylic acid, characterized in that the pH of the starting material is adjusted to a value of 4.45 to 4.85 and at a temperature of + 16 to + 18 ° C, conc. amount of 23 to
CS876284A 1987-08-28 1987-08-28 A method of isolating plasma proteins from ram plasma or serum CS261849B1 (en)

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