John F Kellie1, Jonathan R Kehler1, Molly Z Karlinsey1, Scott G Summerfield1,2. 1. Bioanalysis, Immunogenicity & Biomarkers, In vitro/In vivo Translation Platform, R&D Platform Technology & Science, GSK, 709 Swedeland Rd. King of Prussia, PA, 19460, USA. 2. Bioanalysis, Immunogenicity & Biomarkers, In vitro/In vivo Translation Platform, R&D Platform Technology & Science, GSK, David Jack Centre for R&D, Park Road, Ware, Hertfordshire SG12 0DP, UK.
Abstract
AIM: Typically, quantitation of biotherapeutics from biological matrices by LC-MS is based on a surrogate peptide approach to determine molecule concentration. Recent efforts have focused on quantitation of the intact protein molecules or larger mass subunits of monoclonal antibodies. To date, there has been limited guidance for large or intact protein mass quantitation for quantitative bioanalysis. METHODOLOGY: Intact- and subunit-level analyses of biotherapeutics from biological matrices are performed at 12-25 kDa mass range with quantitation data presented. RESULTS: Linearity, bias and other metrics are presented along with recommendations made on the viability of existing quantitation approaches. CONCLUSION: This communication is intended to start a discussion around intact protein data analysis and processing, recognizing that other published contributions will be required.
AIM: Typically, quantitation of biotherapeutics from biological matrices by LC-MS is based on a surrogate peptide approach to determine molecule concentration. Recent efforts have focused on quantitation of the intact protein molecules or larger mass subunits of monoclonal antibodies. To date, there has been limited guidance for large or intact protein mass quantitation for quantitative bioanalysis. METHODOLOGY: Intact- and subunit-level analyses of biotherapeutics from biological matrices are performed at 12-25 kDa mass range with quantitation data presented. RESULTS: Linearity, bias and other metrics are presented along with recommendations made on the viability of existing quantitation approaches. CONCLUSION: This communication is intended to start a discussion around intact protein data analysis and processing, recognizing that other published contributions will be required.
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