论文部分内容阅读
目的:测定重组人组织型纤溶酶原激活剂改构体(combinant human mutant-threonine mutant-asparagine mutant-lysine tissue type plasminogen activator,rhTNK-tPA)单链含量。方法:将样品用二硫苏糖醇(DTT)打开二硫键,采用分子排阻色谱(size-exclusion high performance liquid chromatograph,SEC-HPLC)法测定。色谱柱:Tsk-Gel2000SWxl;流动相:0.2 mol·L-1PB-0.1%SDS水溶液;流速:0.5 mL·min-1;检测波长:214 nm;柱温:23℃;检测时间:30 min。结果:试验精密度和重复性良好;分离度不低于1.1;3批rhTNK-tPA原液的单链含量分别为74.41%、75.39%、74.43%,3批成品的单链含量分别为73.82%、74.76%、74.91%。结论:本方法经方法学验证可用于rhTNK-tPA产品的常规检定。
Objective: To determine the single-stranded content of recombinant human mutant-threonine mutant-asparagine mutant-lysine tissue type plasminogen activator (rhTNK-tPA). Methods: Disulfide bonds were opened with dithiothreitol (DTT) and determined by size-exclusion high performance liquid chromatography (SEC-HPLC). Column: Tsk-Gel2000SWxl; mobile phase: 0.2 mol·L-1 PB-0.1% SDS in water; flow rate: 0.5 mL · min-1; detection wavelength: 214 nm; Results: The precision and repeatability of the assay were good and the resolution was not less than 1.1. The single-stranded contents of the three batches of rhTNK-tPA were 74.41%, 75.39% and 74.43% respectively. The single-stranded contents of the three batches were 73.82% 74.76%, 74.91%. Conclusion: This method is validated by methodology for the routine assay of rhTNK-tPA products.