分析测试学报2017，Vol.36Issue(3)：343-348,6.DOI:10.3969/j.issn.1004-4957.2017.03.008
全柱成像毛细管等电聚焦电泳分析蛋白质药物电荷异质性
Analysis of Peptide and Protein Drugs Isoelectric Point by Full Column Imaging cIEF-WCID
摘要
Abstract
The application effect of isoelectric points(pI) of peptide and protein drugs containing different molecular weights and complex charge heterogeneity was evaluated by capillary isoelectric focusing electrophoresis-whole column imaging detection(cIEF-WCID).The pI of standard polypeptide was mensured repeatedly to verify that cIEF-WCID had a high accuracy and a good repeatability (RSD ＜ 0.50％).Four serum protein isomers had achieved baseline separation.Gan Lai insulin was detected only a single characteristic peaks(pI 5.95 ±0.01) when repeating the test.The proportion differences of the characteristic peak'PI were siguificant,when comparing the recombinant human growth hormone bulk and final,and the final appeared new characteristic peaks.Comparing the imported and domestic Avastin,manufacturer 1 and 3 were found to be consistent with the innovator drug,and the migration behavior of main component in manufacturer 1 was highly consistent with the innovator drug,however,the charge heterogeneity of manufacturer 2 was influenced because of lose K,N-term pyro-Glu.The optimized pI detecting conditions of the recombinant human follicle stimulating hormone were obtained by studying co-solvent concentration,electrolyte and focusing time,namely Co-solvent:2 mol/L urea,ampholytes at pH 2.5-5.0 and pH 3.0-10.0 mixing by 1 ∶ 1,and focus voltage:1 000 V(1 min) to 1 800 V(4 min) and to 2 200 V(1 min).cIEF-WCID was fast and accurate,and it owned excellent resolution and reproducibility,especially,it could trace the focusing process of the analytes.Therefore,it was fit for analysing complex charge heterogeneity of protein drugs.关键词
全柱成像毛细管等电聚焦电泳/电荷异质性/重组蛋白质药物/抗体/药学研究Key words
capillary isoelectric focusing electrophoresis-whole column imaging detection (cIEF-WCID)/charge heterogeneity/recombinant protein drug/antibody/pharmaceutical research分类
化学化工引用本文复制引用
孟晓光,甄里,刘悦玫,侯利平,刘丹,李斌,欧阳伟民,魏开华..全柱成像毛细管等电聚焦电泳分析蛋白质药物电荷异质性[J].分析测试学报,2017,36(3):343-348,6.基金项目
国家高技术研究发展计划863项目(2014AA020906) （2014AA020906）
国家科技部重大仪器专项(2012YQ18011710) （2012YQ18011710）
蛋白质组学国家重点实验室课题(SKLP-0201412) （SKLP-0201412）
