| 重组单抗药物的肽图分析 |
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| 引用本文： | 魏敬双,程立均,刘进怀,贾茜. 重组单抗药物的肽图分析[J]. 生物技术通报, 2008, 0(Z1) |
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| 作者姓名： | 魏敬双 程立均 刘进怀 贾茜 |
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| 作者单位： | 华北制药集团新药研究开发有限责任公司,石家庄,050015 |
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| 基金项目： | “十一五”863计划生物和医药技术领域重大项目基金资助(2006AA02A247) |
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| 摘 要： | 建立了重组单抗药物的肽图分析方法。在变性条件下向抗体溶液加入还原剂,打开抗体内部所有交联的二硫键,再加入烷基化试剂封闭所有的自由巯基,使抗体分子在溶液中以游离伸展肽链的形式存在。加入胰蛋白酶,将充分伸展的肽链酶解成小的肽段。用反相高效液相色谱层析分析肽图谱。对连续3批中试产品及理化测定对照品进行肽图分析,各样品均能酶解完全,批间肽图谱一致。该方法实用有效,适于进行重组单抗等结构复杂的大分子蛋白药物的肽图检查分析。
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| 关 键 词： | 重组单抗 肽图 抗狂犬病毒单抗 胰蛋白酶 反相HPLC |
Peptide Mapping of Recombinant Monoclonal Antibody Pharmaceuticals |
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| Wei Jingshuang,Cheng Lijun,Liu Jinhuai,Jia Qian. Peptide Mapping of Recombinant Monoclonal Antibody Pharmaceuticals[J]. Biotechnology Bulletin, 2008, 0(Z1) |
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| Authors: | Wei Jingshuang Cheng Lijun Liu Jinhuai Jia Qian |
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| Affiliation: | New Drug R&D Center;North China Pharmaceutical Corporation;Shijiazhuang 050015 |
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| Abstract: | A method was established for peptide mapping of recombinant monoclonal antibody pharmaceuticals.In denaturing conditions,reducing reagent was added to recombinant antibody solution,all the crosslinked disulfide bonds were broken,and then alkylation reagent was added to block the free sulfhydryl residues,the resultant antibody molecules stayed in the solution as dissociated and extensively stretched peptide chains.The trypsin was added to the solution and the antibody was digested to small peptide fragments.... |
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| Keywords: | Recombinant monoclonal antibody Peptide mapping Anti-rabies monoclonal antibody Trypsin Reversed phase HPLC |
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