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目的:建立蚓激酶的纤溶酶谱鉴别方法,同时考察5个企业产品及同一企业不同批次的一致性。方法:纤溶酶谱法,纤维蛋白原终浓度为5×10~(-4)g·ml~(-1),2.5%Triton-x-100溶液中复性30 min,在PBS缓冲液(p H=7.4)中37℃孵育30 min,供试品蛋白质浓度为5.0~37.5μg。结果:该方法灵敏度高,5家企业的活性蛋白条带均分布在15~40KD间,有6个相同特征活性蛋白条带;5个企业产品之间酶谱图略有差异,同一企业不同批次产品批间无差异。结论:该方法专属性较强,能够反映蚓激酶活性成分的分子量大小分布及种类,也能反映产品的批间一致性,生产工艺稳定性,且易于标准化,可作为蚓激酶的鉴别方法,同时为评价上市后产品质量一致性奠定了基础。
OBJECTIVE: To establish a fibrinolytic method for the identification of lumbrokinase, and to examine the consistency of five products and different batches of the same enterprise. Methods: Fibrinolysis was used to determine the final fibrinogen concentration of 5 × 10 ~ (-4) g · ml ~ (-1). After refolding in 2.5% Triton-x-100 solution for 30 min, p H = 7.4) for 30 min at 37 ° C for a protein concentration of 5.0 to 37.5 μg. Results: The sensitivity of the method was high. The active protein bands of the five companies were all distributed between 15 and 40 kD, and there were six bands with the same characteristic active protein. The zymogram of the five enterprises was slightly different, There is no difference between the second product batch. CONCLUSION: This method is highly specific and can reflect the molecular size distribution and species of active ingredients of lumbrokinase. It can also reflect the consistency between lots and the stability of the production process, and is easy to standardize and can be used as the identification method of lumbrokinase. Meanwhile, To evaluate the consistency of product quality after listing laid the foundation.