目的 以健康人血浆为原料，探索人凝血酶原的激活条件，提升产品比活性和得率。方法 采用DEAE sephadex A-50凝胶吸附分离健康人合并血浆制备人凝血酶原，并考察Ca²⁺浓度、初始人凝血酶原浓度、激活温度、初始pH以及激活时间对人凝血酶原激活效果的影响；采用建立的激活条件，经人凝血酶原纯化、人凝血酶原激活、阳离子交换层析等步骤，制备3批实验成品并完成质量指标检测。结果 DEAE sephadex A-50凝胶吸附步骤人凝血酶原平均回收率为81.98%。在氯化钙质量浓度为0.6%、初始人凝血酶原浓度为10 国际单位（international unit，IU）/mL、温度2~8 ℃、初始pH7.45条件下，放孵12 d，每单位人凝血酶原激活可以得到209.50 IU的人凝血酶。3个实验批次阳离子交换层析步骤人凝血酶平均回收率为77.57%，人凝血酶制品比活性均值为1 078.06 IU/mg，理论每吨血浆最高可以制备约6.40×10⁷ IU的人凝血酶产品，3批成品均符合国家药典标准。结论 建立的激活条件可高效激活人凝血酶原，经过阳离子交换层析制备的人凝血酶产品比活性高、回收率高，可大幅降低生产成本，适用于规模化生产。

Objective To explore the activation conditions of human prothrombin with healthy human plasma as raw material, and to improve the specific activity and yield of the product.Methods DEAE sephadex A-50 gel adsorption method was used to separate healthy human plasma mixture to prepare human prothrombin. Five factors including Ca²⁺ concentration, initial human prothrombin concentration, activation temperature, initial pH and activation duration were investigated to optimize the activation conditions of human prothrombin. Using the established activation conditions, 3 batches of final products were produced through purification of human prothrombin, activation of human prothrombin, cation exchange chromatography and other steps, followed by comprehensive quality testing.Results The average yield of human prothrombin in the DEAE sephadex A-50 gel adsorption step was 81.98%. Under the conditions of CaCl₂ mass concentration 0.6%, initial human prothrombin concentration 10 international unit (IU)/mL, temperature 2-8 ℃, initial pH7.45 and incubation for 12 d, 209.50 IU of human thrombin was obtained per unit of human prothrombin. The average recovery rate of human thrombin in the cation exchange chromatography step of the final products was 77.57%, and the average specific activity of human thrombin products was 1 078.06 IU/mg. Theoretically, about 6.40×10⁷ IU of human thrombin product could be prepared per ton of plasma, and all three batches of final products met the Chinese pharmacopoeia standards.Conclusion The established activation conditions can efficiently activate human prothrombin, and the human thrombin product prepared by cation exchange chromatography has high specific activity and high yield, which can greatly reduce the production cost, and is suitable for large-scale production.