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目的 :用一种新的离子交换层析法从人血浆冷沉淀中制备高纯FVIII。方法 :溶解后冷沉淀经氢氧化铝吸附 ,低温下酸沉淀和聚乙二醇沉淀后用 0 .3%磷酸三正丁酯和 1%Tween 80处理以灭活脂包膜病毒 ,然后过DEAE FractogelTSK 6 5 0M层析柱。绝大部分杂蛋白和磷酸三正丁酯 Tween 80直接流穿 ,而吸附的FVIII凝集活性蛋白通过增加离子强度洗脱 ,并直接除菌分装而无需超滤或加入蛋白稳定剂。结果 :层析收率达 70 %～ 90 % ,制品特异性活性超过 10 0IU·mg-1,相对血浆纯化倍数超过 5 0 0 0倍。结论 :该制剂纯度高于传统FVIII制剂 ,应用于甲型血友患者的治疗具有更好的耐受性和效果
OBJECTIVE: To prepare high purity FVIII from human plasma cryoprecipitate by a new ion exchange chromatography. METHODS: The cryoprecipitate was adsorbed onto aluminum hydroxide, and after acid precipitation and polyethylene glycol precipitation at low temperature, it was treated with 0.3% tri-n-butyl phosphate and 1% Tween 80 to inactivate the lipid enveloped virus and then subjected to DEAE FractogelTSK 6 50M column. Most of the hybrid proteins and tri-n-butyl phosphate Tween 80 flow through directly, while the adsorbed FVIII agglutination active protein elutes by increasing ionic strength and is directly sterilized and dispensed without ultrafiltration or addition of protein stabilizers. Results: The chromatographic yield was 70% -90%. The specific activity of the product was over 100 IU · mg-1, and the relative plasma purification times exceeded 500 times. Conclusion: The purity of the preparation is higher than the traditional FVIII preparation and has better tolerability and efficacy in the treatment of hemophilia A patients