微柱高效液相色谱分析基因工程药物rhG-CSF肽谱的应用研究

Study on peptide mapping analysis of recombinant medicine rhG-CSF with microcolumn HPLC

用二硫代苏糖醇还原基因工程药物产品中的二硫键，并用碘乙酰胺封闭巯基，再用胰蛋白酶裂解，经微柱高效液相色谱分离分析rhG－CSF产品的肽谱，3个批号批间结果完全一致。该方法具有系统操作误差小，用样品量少，较常现高效液相色谱检测灵敏度高，重视性好的优点。是适用于基因工程药物产品肽谱分析，衡量产品生产纯化工艺稳定性比较优秀的方法之一。

After the disulfide bond in recombinant medicine, rhG-CSF was reduced by DTT and the sulfydryl group was blocked by iodoacetamide,the rhG-CSF was digested by trypsin and peptide mapping was analyzed by microcolumn-HPLC.Difference in one or two amino acid residues in the peptide of rhG-CSF could be identified. The method shows good sensitivity and reproductivity,and it is suitable for the routine peptide mapping of recombinant medicines.