Rapid assessment of oxidation via middle-down LCMS correlates with methionine side-chain solvent-accessible surface area for 121 clinical stage monoclonal antibodies.

Autor: Yang R; a Protein Analytics, Adimab , Lebanon , NH , USA., Jain T; b Computational Biology, Adimab , Palo Alto , CA , USA., Lynaugh H; a Protein Analytics, Adimab , Lebanon , NH , USA., Nobrega RP; a Protein Analytics, Adimab , Lebanon , NH , USA., Lu X; a Protein Analytics, Adimab , Lebanon , NH , USA., Boland T; b Computational Biology, Adimab , Palo Alto , CA , USA., Burnina I; a Protein Analytics, Adimab , Lebanon , NH , USA., Sun T; a Protein Analytics, Adimab , Lebanon , NH , USA., Caffry I; a Protein Analytics, Adimab , Lebanon , NH , USA., Brown M; a Protein Analytics, Adimab , Lebanon , NH , USA., Zhi X; a Protein Analytics, Adimab , Lebanon , NH , USA., Lilov A; a Protein Analytics, Adimab , Lebanon , NH , USA., Xu Y; a Protein Analytics, Adimab , Lebanon , NH , USA.
Jazyk: angličtina
Zdroj: MAbs [MAbs] 2017 May/Jun; Vol. 9 (4), pp. 646-653. Date of Electronic Publication: 2017 Feb 14.
DOI: 10.1080/19420862.2017.1290753
Abstrakt: 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% H 2 O 2 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: MEDLINE
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