Rapid assessment of oxidation via middle-down LCMS correlates with methionine side-chain solvent-accessible surface area for 121 clinical stage monoclonal antibodies
| Autor: | R. Paul Nobrega, Tingwan Sun, Todd Boland, Asparouh Lilov, Michael Brown, Yingda Xu, Xiaoyong Zhi, Rong Yang, Isabelle Caffry, Irina Burnina, Xiaojun Lu, Heather Lynaugh, Tushar Jain |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Models Molecular medicine.drug_class Immunology Cleavage (embryo) Monoclonal antibody Immunoglobulin light chain Mass Spectrometry Accessible surface area 03 medical and health sciences chemistry.chemical_compound Methionine Side chain medicine Immunology and Allergy Humans Chromatography Antibodies Monoclonal Hydrogen Peroxide Combinatorial chemistry 030104 developmental biology HEK293 Cells chemistry Forced degradation Chemical stability Oxidation-Reduction Chromatography Liquid Reports |
| Zdroj: | mAbs. 9(4) |
| ISSN: | 1942-0870 |
| Popis: | Susceptibility of methionine to oxidation is an important concern for chemical stability during the development of a monoclonal antibody (mAb) therapeutic. To minimize downstream risks, leading candidates are usually screened under forced oxidation conditions to identify oxidation-labile molecules. Here we report results of forced oxidation on a large set of in-house expressed and purified mAbs with variable region sequences corresponding to 121 clinical stage mAbs. These mAb samples were treated with 0.1% H2O2 for 24 hours before enzymatic cleavage below the hinge, followed by reduction of inter-chain disulfide bonds for the detection of the light chain, Fab portion of heavy chain (Fd) and Fc by liquid chromatography-mass spectrometry. This high-throughput, middle-down approach allows detection of oxidation site(s) at the resolution of 3 distinct segments. The experimental oxidation data correlates well with theoretical predictions based on the solvent-accessible surface area of the methionine side-chains within these segments. These results validate the use of upstream computational modeling to predict mAb oxidation susceptibility at the sequence level. |
| Databáze: | OpenAIRE |
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