A workflow for absolute quantitation of large therapeutic proteins in biological samples at intact level using LC-HRMS. (August 2016)

• Record Type: Journal Article
• Authors: Jian, Wenying; Kang, Lijuan; Burton, Lyle; Weng, Naidong
• Is Part Of: Bioanalysis. Volume 8:Number 16(2016)
• Page Start: 1679
• Page End: 1691
• Publication Date: 2016-08
• Subjects: bioanalysis -- intact level -- LC-HRMS -- protein bioanalysis -- quantitation; Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio-2016-0096
• Languages: English
• ISSNs: 1757-6180

Abstract

Aim: The commonly used LC–MS workflow to quantify protein therapeutics in biological samples is 'bottom-up' approach. In this study, the aim is to establish 'top-down' approach for absolute quantitation of therapeutic antibodies or proteins of similar sizes in biological samples at intact level.Materials & methods : Using a recombinant human monoclonal antibody as the model molecule, we present a workflow to measure large therapeutic proteins in plasma at intact level based on deconvoluted high-resolution MS (HRMS) peaks. A novel MultiQuant™ software function was developed to automatically deconvolute the peaks and process the data.Results & conclusion: The workflow showed satisfying performance. This is a proof of concept study demonstrating the feasibility of bioanalysis of large therapeutic proteins at intact level using LC-HRMS.