Study on the separation and purification of degradation fragment of human serum albumin

OBJECTIVE To study on the separation and purification of degradation fragment of human serum albumin(HSA) in the chromatography column,and to optimize the process condition.METHODS Firstly,Sephadex G-100 and Superdex 75 were used respectively to separate and purify the degradation products of HSA.Secondly,the carboxymethylcellulose(CM-Sepharose FF) and DEAE-cellulose(DEAE-Sepharose FF) were used respectively under the different pH of phosphate buffered saline for further purification.The process condition was optimized by the resolution and protein recovery.RESULTS The separating effect of Superdex 75 was better than that of Sephadex G-100,and the degradation products were separated into two parts Ⅰ and Ⅱ,and protein recovery was 61.82%±2.01%.CM-Sepharose FF was superior to DEAE-Sepharose FF,part Ⅰwas fractionated into HSA polypeptide fragment A 299-585 and protein recovery was 62.07%±1.12%.Part Ⅱ was fractionated into HSA polypeptide fragment A 1-123 and A 124-298,and protein recovery was 66.29%±1.79%.CONCLUSION It is experimentally shown that high resolution and protein recovery of the degradation products of HSA are obtained in fixed bed chromatography with Superdex 75 and CM-Sepharose FF.