重组人C1酯酶抑制剂中聚合物含量的测定

目的：建立分析重组人C1酯酶抑制剂（rhC1INH）中聚合物含量的分子排阻高效液相色谱（SEC-HPLC）方法。方法：rhC1INH稀释后，采用TSK GEL3000SW色谱柱，214 nm检测波长对其聚合物进行检测，以250 mmol·L^-1磷酸缓冲液（pH 6.8）为流动相，流速0.5 mL·min^-1进行洗脱。结果：在25.0~85.0 μg·mL^-1浓度范围内，rhC1INH标准品的峰面积与浓度的线性关系良好（R²平均值为0.997）。rhC1INH样品中分离到3个聚合物峰，采用面积归一法计算6次进样的聚合物的平均含量分别为0.11%、0.15%和0.60%，RSD分别为10.7%、9.0%和3.9%。4批次rhC1INH样品中总聚合物的含量分别为0.82%、1.01%、0.97%和0.92%。结论：SEC-HPLC方法适用于rhC1INH中聚合物的检测，为其聚合物质量标准的建立和产品评价奠定了基础。

Objective: To develop a size-exclusion high performance liquid chromatograph(SEC-HPLC) for determination of the aggregates protein in recombinant human C1 esterase inhibitor(rhC1INH).Methods: The rhC1INH samples were diluted with mobile phase and separated on a column of TSK GEL3000SW column.The mobile phase was 250 mmol·L^-1 phosphate buffer(pH 6.8),the flow rate was 0.5 mL·min^-1,and detection wavelength was 214 nm.Results: The peak area of rhC1INH standard showed a good liner relationship with the concentration over the range of 25.0-85.0 μg·mL^-1(the mean R²=0.997).Three aggregate peaks were isolated from rhC1INH, and the mean contents of three peaks were 0.11%,0.15% and 0.60%,with RSD of 10.7%,9.0% and 3.9%,respectively.The total aggregates in four batches of rhC1INH were 0.82%,1.01%,0.97% and 0.92%,respectively.Conclusion: The SEC-HPLC revealed a feasible detection for the rhC1INH aggregates,and provided basis for establishment of aggregates-related quality standards and product evaluation.