重组人促卵泡激素（rhFSH）制剂的SDS-PAGE/Western_blot鉴别方法研究___

目的：建立可用于不同剂型重组人促卵泡激素（rhFSH）产品质量控制的特异性SDS-PAGE/Western blot鉴别方法。方法：用SDS-PAGE电泳分离由不同生产企业生产的不同剂型rhFSH产品，通过半干电泳将凝胶上的待测蛋白转移至硝酸纤维素膜（NC膜），采用特异性抗体在NC膜上进行免疫杂交。通过检测灵敏度、线性和专属性试验，进行方法学考察。结果：对rhFSH特异性检测灵敏度可达5 ng；对3个厂家生产的3种注射液、1种注射用冻干制剂检测，均可得到阳性结果，且试验结果不受不同剂型中的防腐剂、稳定剂和其他辅料干扰；用本法对等量的其他重组糖蛋白激素如重组人促黄体激素（rhLH）、重组人绒促性素（rhCG）测定结果均为阴性。结论：本方法灵敏度高、专属性强，可作为rhFSH的特异性鉴别方法，用于不同剂型rhFSH产品的质量控制。

Objective:To establish the method of specific identification by SDS-PAGE/Western blot for the different dosage forms of the recombinant human follicle stimulating (rhFSH).Methods:The samples of different dosage forms of the rhFSH preparations produced by different manufactures were subjected to SDS-PAGE under non-reducing conditions.After transferring the proteins in lanes from the gel to the nitrocellulose filter (NC) by half-dry electrophoresis,the specific antibody to FSH was used to immunodetect the rhFSH on the NC.The method was evaluated by sensitivity determination,linear study and specificity test.Results:A minimum of 5 ng of the rhFSH could be detected in the sensitivity test.All the results obtained from the samples of three kinds of injection solution and one kind of power for injection which produced by three different manufacturers were positive.Furthermore,these results would not be influenced by preservatives,stabilizers and other excipients formulated in different dosage forms.All the results were negative when the other glycoprotein hormones such as the recombinant human luteum stimulating (rhLH) and the recombinant human chorionic gonadotrophin (rhCG) were tested by the method described above.Conclusion:The method developed here has high sensitivity and high specificity.It may be served as the routine method of specific identification in quality control for different dosage forms of the rhFSH products.