A workflow for absolute quantitation of large therapeutic proteins in biological samples at intact level using
Wenying Jian1, Lijuan Kang1, Lyle Burton2
1Pharmacokinetics, Dynamics, & Metabolism (PDM), Janssen Research & Development, Johnson & Johnson, 1400 McKean Road, Spring House, PA 19477, USA.
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.
