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Analysis of Peptide and Protein Drugs Isoelectric Point by Full Column Imaging cIEF-WCID |
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DOI: |
KeyWord:capillary isoelectric focusing electrophoresis-whole column imaging detection(cIEF-WCID) charge heterogeneity recombinant protein drug antibody pharmaceutical research |
Author | Institution |
MENG Xiao-guang,ZHEN Li,LIU Yue-mei,HOU Li-ping,LIU Dan,LI Bin,OUYANG Wei-min,WEI Kai-hua |
1.北京正旦国际科技有限责任公司;2.北京蛋白质组研究中心,蛋白质组学国家重点实验室,国家蛋白质科学中心(北京);3.江汉大学护理与医学技术学院;4.北京绿绵科技有限公司;5.军事医学科学院放射与辐射医学研究所 |
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Hits: 2012 |
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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 peakPI were significant,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 . |
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