2020
DOI: 10.1021/jasms.0c00036
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Interlaboratory Study for Characterizing Monoclonal Antibodies by Top-Down and Middle-Down Mass Spectrometry

Abstract: The Consortium for Top-Down Proteomics (www.topdownproteomics.org) launched the present study to assess the current state of top-down mass spectrometry (TD MS) and middle-down mass spectrometry (MD MS) for characterizing monoclonal antibody (mAb) primary structures, including their modifications. To meet the needs of the rapidly growing therapeutic antibody market, it is important to develop analytical strategies to characterize the heterogeneity of a therapeutic product's primary structure accurately and r… Show more

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Cited by 83 publications
(86 citation statements)
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“…MALDI-ISD FT-ICR MS has previously been explored for protein characterization, where it likewise showed vast complementarity to other fragmentation techniques. 48 , 49 , 66 …”
Section: Resultsmentioning
confidence: 99%
“…MALDI-ISD FT-ICR MS has previously been explored for protein characterization, where it likewise showed vast complementarity to other fragmentation techniques. 48 , 49 , 66 …”
Section: Resultsmentioning
confidence: 99%
“…High-resolution mass spectrometry (HRMS) was employed to confirm the change in glycosylation status between wild type and mutated variants. In order to reduce complexity at the intact mass level a “middle-down” approach was utilized (55, 56). Intact antibodies were incubated with IdeS protease, followed by reduction of disulfide bonds.…”
Section: Methodsmentioning
confidence: 99%
“…Purification of complex protein mixture is especially challenging in biological samples. Top-down approaches can be helpful to evaluate the total modification status on an individual protein [ [93] , [94] , [95] , [96] ]. Small modifications with low abundancies require more sophisticated, high-resolution instruments such as Q-TOFs, Orbitraps or FT-ICR MS. Another option to explore minor modifications occurring on a small percentage of proteins is fragmentation of intact proteins.…”
Section: Analysis Of Oxidative Protein Modificationsmentioning
confidence: 99%