Comparison of four pre-column treatment methods for human plasma coagulation factorⅧpre-purification / 军事医学

ABSTRACT

Objective To compare four pretreatment methods of human plasma coagulation factor Ⅷ( FⅧ) before column purification.Methods Cryoprecipitate was dissolved in Tris , before the supernatant was treated with glycine precipitation, PEG precipitation, acid precipitation and aluminum hydroxide gel adsorption , respectively.Activated partial thromboplastin (APTT) was used to measure the activity of the supernatant clotting FⅧ after treatment.The total activity recovery and specific activity of the final samples were used to weigh the efficacy of those methods .The purity of the intend-ed protein was estimated by non-reducing SDS-PAGE electrophoresis .Results Total activity recovery of glycine precipitati-on was the highest (94.00%±7.60%), followed by that of acidic precipitation (89.47%±2.60%) and PEG precipita-tion (80.92%±9.67%) methods.The lowest was aluminum hydroxide gel adsorption (78.65%±7.52%).Glycine precipitation and PEG precipitation could more effectively remove contaminating protein than acid precipitation and aluminum hydroxide gel adsorption .Treated by four different methods , the specific activity of FⅧ of glycine precipitation sample was the highest (0.6856 ±0.1258 IU/mg), followed by PEG precipitation (0.5773 ±0.0787 IU/mg) and acidic precipitation (0.3885 ±0.0301 IU/mg).The specific activity of aluminum hydroxide gel adsorption was the lowest (0.2879 ±0.0472 IU/mg).Conclusion PEG precipitation is more effective for the actual production process than the other three methods .