Top-down LC–MS quantitation of intact denatured and native monoclonal antibodies in biological samples. (28th June 2018)

• Record Type: Journal Article
• Authors: Zhang, Liyun; Vasicek, Lisa A; Hsieh, SuChun; Zhang, Shuli; Bateman, Kevin P; Henion, Jack
• Is Part Of: Bioanalysis. Volume 10:Number 13(2018)
• Page Start: 1039
• Page End: 1054
• Publication Date: 2018-06-28
• Subjects: antibody -- biotherapeutic quantitation -- denatured and native protein conformation -- immunocapture -- intact top-down protein quantitation -- multidimensional LC–MS -- non-specific binding -- NSB -- top-down QTOF LC/MS; Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio-2017-0282
• Languages: English
• ISSNs: 1757-6180

Abstract

Aim: The requirements for developing antibody biotherapeutics benefit from understanding the nature and relevant aspects of the entire molecule. The method presented herein employs on-line multidimensional LC–quadrupole time-of-flight (QTOF)-MS for the quantitative determination of an antibody isolated from biological samples while maintaining the intact native biologically active conformation of the antibody.Results: Following method optimization for a model antibody, an incurred biotherapeutic in cynomologus monkey was quantified in its intact top-down native conformation. A partial method validation demonstrated acceptable precision and accuracy although improved sensitivity requires further studies.Conclusion: An on-line multidimensional LC–MS approach presents a proof-of-principle example for quantifying an intact, native antibody isolated from an incurred biological sample via immunoaffinity techniques coupled with top-down QTOF LC–MS bioanalysis.