A Rapid, Automated Immunoaffinity LC-MS/MS Workflow

Part # PS320

Abstract

Michael Rosenblatt¹, Gregory Barrett-Wilt², Nidhi Nath¹, Lyndsey Jager¹, Marjeta Urh^1
1Promega Corporation, 2800 Woods Hollow Rd, Madison, WI 53711 ; ²University of Wisconsin, 425 Henry Mall Madison WI 53706

The analysis of Biotherapeutic antibody metabolism in various animal models is a critical requirement during drug development. As the number of potential antibody targets have increased targets as well as Biosimilar development, there is an immediate need to increase sample throughput. Toward this end, we have developed a streamlined workflow that involves mAb enrichment, using highcapacity magnetic streptavidin particles, followed by proteolysis using a commercially available Rapid Digestion kit. The rapid enrichment/digestion step combined with direct LC-MS/MS analysis (no offline cleanup required) allows for protocols that can be carried out in very short time periods (as short as 2–3 hours). The protocol is very simple, requires routine laboratory equipment, and can be easily automated.

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