Parameters optimization in Heparin affinity chromatography purification of human coagulation factor Ⅸ
10.13303/j.cjbt.issn.1004-549x.2021.01.008
- VernacularTitle:响应面法优化人凝血因子Ⅸ肝素亲和层析条件研究
- Author:
Chuan HU
1
;
Guanwen LUO
1
;
Wenshan LI
1
;
Changwen YU
1
;
Bobo YANG
1
Author Information
1. Shuanglin Biological Pharmaceutical Co LTD, Zhanjiang 524000, China
- Publication Type:Journal Article
- Keywords:
human coagulation factor Ⅸ;
heparin affinity chromatography;
response surface method;
process optimization
- From:
Chinese Journal of Blood Transfusion
2021;34(1):26-29
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To optimize the purification conditions of heparin affinity chromatography in the purification of human coagulation factor Ⅸ by response surface method and establish the optimal chromatography process parameters. 【Methods】 The effect of sample loading temperature on purification efficiency was analyzed through single factor test. Three-factor three-level response surface method was used to optimize the chromatographic elution conditions. The Folin phenol method and the automatic hemagglutination analyzer were used to determine the total protein content and human coagulation factor Ⅸ titer, respectively. The purification effect was evaluated by activity index and process recovery rate. 【Results】 The optimized optimal chromatographic conditions were loading at 5 ℃, washing 4 CV, eluent formulation of sodium citrate 20 mmol/L, arginine hydrochloride 18.7 mmol/L, NaCl 611.6 mmol/L and pH 7.5; under this optimal setting, the recovery rate of the chromatographic process was (46.6±2.9) %, titer of factor Ⅸ rated to (68.4±4.7) IU/mL and specific activity was (62.8±3.3) IU/mg. 【Conclusion】 The optimized parameters of heparin affinity chromatography process by response surface method can produce better purification effect on human coagulation factor Ⅸ.
