CS258203B1 - A method for isolating animal albumin - Google Patents

Abstract

The solution can be used in the immunological-biological production of veterinary medicinal products. The aim is to introduce into production a new technology for the isolation of animal albumin, in which the native nature of the protein is preserved and a higher yield is achieved compared to the current state of knowledge. The isolation procedure presented represents only one operational step and can be performed at room temperature. The isolation procedure is simplified by stabilizing the blood plasma or serum with N-acetyl-DL-tryptophan and Na-caprylate, adjusting the pH to 6.0 to 6.1 and adding 96% ethanol in an amount of 12 to 18 volume percent. By subsequently heating the solution at 45 to 55 °C and adjusting the pH to 5.0 to 5.1, the precipitate formed is removed and the supernatant represents albumin with at least 95% electrophoretic purity.

Description

258203 2

The present invention relates to a method for isolating animal albumin.

Up to now, on an industrial scale, animal plasma albumin or serum albumin has been isolated either by extraction with suitable salts or fractions of organic solvents, especially ethanol (CohnE. J. et al., J. Amer. Chem. Soc., 68, 459 (1946); Nitschmann, H. et al. al .: Helv Chim Acta, 37, 866 (1954)). The disadvantage of the mentioned technological processes is a long technological cycle, energy demands and demanding technological equipment. Also known are thermal denaturation methods for albumin isolation (Hoch H., Chanutin: Arch. Of Biochem. And Biophys., 51, 27, 1954; Lundblad JI: Vox sang., 5,122, 1971; Schneider W. et al .: Folia haematol., 104, 116, 1977).

However, the disadvantage of these is that the ballast proteins are denatured at high temperatures (+70 ° C), which adversely affects the nativity and yield of albumin. Also, the isolation of albumin in the production of Plazbion starts from the supernatant after separation of the immunoglobulins, it also uses a relatively high denaturation temperature (60 ° C) and a long denaturation of the ballast proteins (two hours) and finally achieves a relatively low electrophoretic purity (max. 80%) and yield of the final product.

The invention solves the isolation of albumin from animal plasma, respectively. serum by a one-stage techno-logical operation in which all non-albumin plasma proteins are denatured. The advantage of the above process is that in addition to the shortening cycle and the lower denaturation temperature, a yield of approximately 30% is achieved.

The nature of animal albumin isolation is that 0.04 M Na-caprylate and 0.04 M N-acetyl-DL-tryptophan is added to the blood plasma or serum. The pH is adjusted to 6.0 to 6.1 with 1 M acetic acid after homogenization. While stirring, 96% ethanol is added dropwise to a concentration of 12 to 18 volume percent. The solution is then heated with stirring at 45 to 55 UC for 10 minutes. The natopH mixture is adjusted to 5.0 to 5.1 and cooled to +18 to +20 ° C. The precipitate formed is centrifuged and the supernatant is filtered and lyophilized. Example 1 A stabilizer was added to 3600 ml of bovine plasma to a final concentration of 0.04 MNa-caprylate and 0.04 M N-acetyl-D1-tryptophan and pH adjusted to pH 6.0 with 1 M acetic acid. 720 ml ethanol was added to the mixture while stirring, and the solution was heated at 45 ° C for 10 minutes. Then the pH of the solution was adjusted to 5.1 with 1 M acetic acid and the solution was cooled to 18 ° C. The precipitate formed is separated by centrifugation. The supernatant was filtered and lyophilized. The weight of the lyophilized product was 72 g, electrophoretic purity 95.8. Example 2 To 3,800 ml of bovine plasma, a stabilizer was added to a final concentration of 0.04 MNa-caprylate and 0.04 M N-acetyl-D1-tryptophan and pH adjusted to 6.1 with 1 M acetic acid. While stirring, 96% ethanol (480 mL) was added and the solution was heated at 55 ° C for 10 minutes. Then the pH of the solution was adjusted to 5.1 with 1 M acetic acid and the resulting solution was cooled to 20 ° C. The precipitate formed was separated by centrifugation. The supernatant was filtered and lyophilized. Lyophilized product weight 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.