毛细管电泳用于测定人绒促性素的解离亚基及其质量控制

Quality control of human chorionic gonadotrophin subunit dissociation

人绒促性素(hCG)是用于治疗不孕症等的常用药物。其由α亚基和β亚基组成,研究发现hCG在生产及储存过程中均可能产生亚基的解离,从而影响其活性。本文旨在建立hCG解离亚基的测定方法,通过考察样品制备缓冲体系、样品制备温度、分离电压、毛细管温度等,建立了毛细管电泳(CE-SDS)定量测定hCG解离亚基的方法。方法验证结果表明,α亚基和β亚基含量具有良好的灵敏度、线性、准确度和精密度,优于传统的凝胶电泳SDS-PAGE。本文所开发的方法适用于不同来源的hCG,并可应用于指导工艺开发及稳定性研究。因此,该方法应作为hCG生物制品的重要质控方法。

Human chorionic gonadotrophin （hCG）, a glycohormone widely used in treatment of infertility, is a heterodimer composed of an alpha- and a beta-subunit. The heterodimer could be dissociated during production and storage with an impact on its bioactivity. A CE-SDS method for quantitative analysis of hCG subunit dissociation was established in this study by optimization of a variety of method conditions including sample preparation buffer compositions, incubation temperature, separation voltage, and capillary temperature. This method was validated for good sensitivity, linearity, precision, and accuracy for both a- and fl-subunit CE-SDS also showed much better precision and accuracy than SDS-PAGE. The method was successfully used in both recombinant hCG （r-hCG） produced by cell culture and hCG （u-hCG） derived from urine. The CE-SDS method was used in the study of hCG development and stability. Therefore, it is an useful tool for the quality control of hCG.