Wenying Jian1, Lijuan Kang1, Lyle Burton2, Naidong Weng1. 1. Pharmacokinetics, Dynamics, & Metabolism (PDM), Janssen Research & Development, Johnson & Johnson, 1400 McKean Road, Spring House, PA 19477, USA. 2. SCIEX, 71 Four Valley Drive, Concord, ON, Canada, L4K 4V8.
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.
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.
Entities:
Keywords:
LC-HRMS; bioanalysis; intact level; protein bioanalysis; quantitation
Authors: Catherine E DelGuidice; Omnia A Ismaiel; William R Mylott; Moucun Yuan; Matthew S Halquist Journal: Anal Bioanal Chem Date: 2022-04-22 Impact factor: 4.142
Authors: Rachel Liuqing Shi; Gang Xiao; Thomas M Dillon; Arnold McAuley; Margaret S Ricci; Pavel V Bondarenko Journal: MAbs Date: 2021 Jan-Dec Impact factor: 5.857