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| Research of N Terminal Sequencing Methods for Monoclonal Antibody Pharmaceuticals Blocked by Pyroglutamic Acid |
| LI Ming-zhu, HAN Wei-dong, Xing Guang-hui, TENG Zhen-lin, XUE Guo-mei, HOU Chen-rui, RUAN Hong-qiang, CHEN Wei |
| Research Center for Proteome Analysis Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences/Shanghai Applied Protein Technology Co.Ltd, Shanghai 200233, China |
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Abstract Objective: Some monoclonal antibody pharmaceuticals with N terminal sequence blocked by pyroglutamic acid can't be sequenced directly by N terminal sequencer which is designed based on Edman degradation principle. Removal of the pyroglutamic acid to realize de novo sequencing of the antibody. Methods: First, using N terminal sequencer to test the light and heavy chains of antibody without treated. Second, confirm the existing of pyroglumatic acid by LC-MSMS. Last, Cutting pyroglumatic acid with pyroglumate aminopeptidase, and then, sequencing the light and heavy chains of antibody with N terminal sequencer. Conclusion: Antibody's light and heavy chains blocked with pyroglumatic acid can be proved by N terminal sequencer and LC-MSMS. And, N terminal sequence of the samples can be proved the same with theoretical one after removal of pyroglumatic acid.
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Received: 21 February 2013
Published: 25 August 2013
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