CS258203B1 - Method of animal albumin insulation - Google Patents

Abstract

The solution can be used in immuno-biological production of veterinary medicinal products. The aim is to introduce a new production Animal albumin isolation technology at which protein nativity is maintained and higher yields compared to current state of knowledge. Said insulating the procedure represents only one operating stage and can be carried out at room temperature. simplification isolation process is achieved by that blood plasma, respectively. the serum stabilizes N-acetyl-DL-tryptophan and Na-caprylate, pH is adjusted to 6.0 to 6.1 and added 96% ethanol in an amount of 12 to 18 vol percent. Subsequent heating of the solution at 45 to 55 ° C and pH adjustment to 5.0 to The precipitate formed is removed and the supernatant is removed represents at least 95% albumin electrophoretic purity.

Description

The invention relates to a process for the isolation of animal albumin.

To date, on an industrial scale, animal albumin from plasma or serum has been isolated either by salting out with suitable salts or a fraction of organic solvents, in particular ethanol (Cohn EJ et al .: J. Amer. Chem. Soc., 68, 459 (1946); Nitschmann H. et al. Helv Chim Acta

37, 866 (1954). The disadvantages of the mentioned technological procedures are long technological cycle, energy intensity and demanding technological equipment. Thermal denaturation methods for albumin isolation are also known (Hoch H., Chanutin: Arch. Of Biochem. And Biophys., 51, 27, 1954; Lundblad JI: Vox sang., 5,122, 1971; Schneider W. et al .: Folia haematol.

1977).

However, the disadvantage of these is that the ballast proteins are denatured at high temperatures (+ 70 ° C), which adversely affects the native and yield of albumin. Also, the isolation of albumin in the production of Plazbion is based on the supernatant after separation of immunoglobulins, it also uses a relatively high denaturation temperature (60 ° C) and a long period of denaturation of the ballast proteins (two hours) and finally achieves a relatively low electrophoretic purity (max.

%) and the yield of the final product.

The invention provides for the isolation of albumin from animal plasma, respectively. sera by a single-step technological procedure in which all plasma proteins except albumin are denatured. The advantage of this process over the hitherto used is that in addition to shortening the production cycle and lower denaturation temperature, the yield is about 30% higher.

The essence of the isolation of animal albumin is that the blood plasma, respectively. 0.04 M Na-caprylate and 0.04 M N-acetyl-DL-tryptophan are added to the serum. After thorough homogenization, the pH was adjusted to 6.0-6.1 with 1M acetic acid. While stirring, 96% ethanol is added dropwise to a concentration of 12-18% by volume. The solution was stirred with continued heating at 45 to 55 C. ^In a period of 10 minutes. The pH of the mixture was adjusted to 5.0-5.1 and cooled to +18 to +20 ° C. The resulting pre-precipitate is centrifuged and the supernatant is filtered and lyophilized.

Example no. 1

To 3800 ml bovine plasma, a stabilizer was added to a final concentration of 0.04 M Na-caprylate and 0.04 M N-acetyl-D1-tryptophan and adjusted to pH 6.0 with 1 M acetic acid.

To the mixture with stirring was added 96% ethanol in an amount of 720 ml and the solution was heated at 45 ° C for 10 minutes. Then the pH of the solution was adjusted to 5.1 with 1M acetic acid and the subsequent solution was cooled to 18 ° C. The resulting pre-precipitate was separated on a centrifuge. The supernatant was filtered and lyophilized. Weight of lyophilized product 72 g, electrophoretic purity 95.8.

Example # 2

To 3800 ml of bovine plasma, a stabilizer was added to a final concentration of 0.04 M Na-caprylate and 0.04 M N-acetyl-D1-tryptophan and the pH adjusted to 6.1 with 1 M acetic acid.

96% Ethanol (480 ml) was added with stirring, and the solution was heated at 55 ° C for 10 minutes. Then the pH of the solution was adjusted to 5.1 with 1M acetic acid and the subsequent solution was cooled to 20 ° C. The resulting pre-precipitate was separated on a centrifuge. The supernatant was filtered and lyophilized. Weight of lyophilized product 68 g, electrophoretic purity 96.1.

Claims (1)

Animal albumin isolation method, characterized in that the animal plasma or serum is stabilized with 0.04 M Na-caprylate and 0.04 M N-acetyl-DL-tryptophan, the pH is adjusted to 6.0-6.1, added 96% ethanol to 12-18% by volume, heat the solution at 45-55 ° C for 10 minutes, then adjust the pH to 5.0-5.1, cool to +18 to +20 ° C , the precipitate formed is centrifuged and the supernatant is filtered and lyophilized.