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| Primary Structural Ananlysis of Reference IgG1 Monoclonal Antibody |
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| DOI： |
| KeyWord:reference antibody molecular weight isoelectric point(pI) charge heterogeneity peptide mapping |
| Author | Institution |
| CUI Xin-ling,HU Zhi-shang,MENG Xiao-guang,QIAN Xiao-hong,ZHU Tao,YING Wan-tao |
1.College of Biotechnology,Tianjin University of Science & Technology;2.State Key Laboratory of Proteomics,Beijing Proteome Research Center,National Center for Protein SciencesBeijing;3.National Institute of Metrology;4.Beijing C&N International Sci-tech Co.,Ltd.;5.CANSINO BIOLOGICS INC.CanSinoBIO |
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| Abstract: |
| A comprehensive structural analysis on reference monoclonal antibody(IgG1) was conducted,which provides a reference for further in depth research of quality and future quality controling.Exactive plus EMR mass spectrometry was used to detect the precise molecular weight(MW) before and after PNGaseF digestion,then calculating the N glycan content.The MW of the reference antibody ranged from 148 285.0 to 149 020.8,which was 145 810.4 after PNGaseF digestion,and the N glycan content ranged from 1.67% to 2.15%.The WCID-cIEF results showed that the isoelectric point(pI) of the antibody ranged from 8.21 to 8.74,in which the pI of the main component was 8.61 and the relative content was 6143%.Then,the charge heterogeneity was detected by ion exchange chromatography.The contents for basic peak,acid peak and main peak were 4.1%,23.9% and 72.0%,respectively,which were consistent with the pI distribution by WCID-cIEF analysis.Furthermore,the glycopeptide distribution and the glycosylated sites were analyzed by peptide mapping before and after PNGaseF digestion,as well as the complementary determinant region(CDR) of the heavy and light chains,and the oxidation sites and oxidation ratio of methionine.The disulfide bonds were also analyzed by peptide mapping before and after antibody reduction.Finally,the glycation of the C-terminal of the antibody was detected.Results showed that there was no glycation occured in the C-terminal of the reference antibody. |
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