Method for preparing human serum albumin containing heat-treatment
View Patent ↗A method for preparing human serum albumin comprises the step of heat-treating a human serum albumin solution containing contaminants at a pH value in the proximity to the isoelectric point of the contaminants.
1. A method for preparing recombinant human serum albumin (HSA) from yeast, comprising:
(1) subjecting HSA-containing fractions to an alkali-treatment at a pH value ranging from 8 to 11, at a temperature ranging from 0 to 60° C. and for a time of at least 15 minutes to convert multimers of recombinant human serum albumin into monomers, followed by
(2) heat-treating at a pH value in the proximity of the isoelectric point of yeast contaminants, at a temperature ranging from 50 to 70° C., at a pH of 5 to 6, and for a time ranging from 30 minutes to 5 hours.
2. The method of claim 1 , wherein the subjecting is conducted at pH 9.0, at room temperature for 5 hours, and the heat-treating is conducted at pH 5.5, at 60° C. for 1 hour.
3. The method of claim 1 , wherein (1) is conducted at a pH of 8.5 to 9.5.
4. The method of claim 1 , wherein (1) is conducted for a time of not less than 3 hours.
5. The method of claim 1 , wherein (1) is conducted at 25° C.
6. The method of claim 1 , wherein (2) is conducted at a pH of 5.5.
7. The method of claim 1 , wherein (2) is conducted at a temperature of 55 to 60° C.
8. The method of claim 1 , wherein (2) is conducted at a temperature of 60° C.
9. The method of claim 1 , wherein (2) is conducted for one hour.
10. A method for preparing recombinant human serum albumin from yeast, comprising:
(1) subjecting a culture broth of human serum albumin-producing yeast to cation exchange column chromatography, wherein the recombinant human serum albumin is adsorbed to and then eluted from the column,
(2) subjecting the eluate to an alkali-treatment at a pH value ranging from 8 to 11, at a temperature ranging from 0 to 60° C. and for a time of at least 15 minutes, to convert the multimers of the recombinant human serum albumin into monomers,
(3) concentrating the recombinant human serum albumin-containing solution by ultrafiltration, and
(4) heat-treating the concentrate at a pH of 5 to 6.
11. The method of claim 10 , wherein (2) is conducted at pH 9.0, at room temperature for 5 hours, and (4) is conducted at pH 5.5, at 60° C. for 1 hour.
12. The method of claim 10 , wherein (2) is conducted at a pH of 8.5 to 9.5.
13. The method of claim 10 , wherein (2) is conducted for a time of not less than 3 hours.
14. The method of claim 10 , wherein (2) is conducted at 25° C.
15. The method of claim 10 , wherein (4) is conducted at a pH of 5.5.
16. The method of claim 10 , wherein (4) is conducted at a temperature of 55 to 60° C.
17. The method of claim 10 , wherein (4) is conducted at a temperature of 60° C.
18. The method of claim 10 , wherein (4) is conducted for one hour.