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Transglutaminase-based antibody-drug conjugation: antibody site-specific mutation and identification
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Zhao-xiong MA1, Yao XU1, Hong ZHAO2, Fu-mou SUN1, Xin-rong ZHANG1, Min WANG1, *, Juan ZHANG1, *
Acta Pharmaceutica Sinica | 2017, 52(3) : 403 - 408
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Acta Pharmaceutica Sinica | 2017, 52(3): 403-408
• ORIGINAL ARTICLES •
Transglutaminase-based antibody-drug conjugation: antibody site-specific mutation and identification
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Zhao-xiong MA1, Yao XU1, Hong ZHAO2, Fu-mou SUN1, Xin-rong ZHANG1, Min WANG1, *, Juan ZHANG1, *
Affiliations
  • 1. College of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
  • 2. Zhejiang Province Hospital of Traditional Chinese Medicine, Hangzhou 310006, China
Published: 2017-03-12 doi: 10.16438/j.0513-4870.2016-0962
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Transglutaminase (TG) posttranslational modification of antibody permits more precisely conjugating. Based on the amino acid sequence of an anti-CD24 antibody (cG7), this article is aimed to generate a deglycosylated cG7 mutant (cG7Q). Firstly, we introduced additional glutamines at position 297 (N297Q) by site-directed mutagenesis, and then transfected the recombinant plasmids into CHO-s cells via electroporation method and screened by Dot blot assay. Subsequently, cG7Q was expressed and purified through Protein A affinity chromatography, further identified by SDS-PAGE electrophoresis and Western blot. Its affinity was detected with surface plasmon resonance and flow cytometry assay, and ADCC effect was determined by lactate dehydrogenase (LDH) release. Eventually, a cG7 mutant, cG7Q was successfully expressed with sequence-specific conjugation sites for further study.

antibody  /  site-specific mutagenesis  /  transglutaminase  /  antibody-drug conjugate
Zhao-xiong MA, Yao XU, Hong ZHAO, Fu-mou SUN, Xin-rong ZHANG, Min WANG, Juan ZHANG. Transglutaminase-based antibody-drug conjugation: antibody site-specific mutation and identification[J]. Acta Pharmaceutica Sinica, 2017 , 52 (3) : 403 -408 . DOI: 10.16438/j.0513-4870.2016-0962
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Article Info
doi: 10.16438/j.0513-4870.2016-0962
  • Receive Date：2016-09-29
  • Online Date：2026-01-14
  • Published：2017-03-12
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History
  • Received：2016-09-29
  • Revised：2016-11-09
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Affiliations
    1. College of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
    2. Zhejiang Province Hospital of Traditional Chinese Medicine, Hangzhou 310006, China
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小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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