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Exploring peptide capture by anti-protein antibodies for LC-MS-based biomarker determination

preprint
revised on 18.10.2018 and posted on 18.10.2018 by Maren Levernæs, Bassem Farhat, Inger Oulie, Sazan S. Abdullah, Elisabeth Paus, Léon Reubsaet, Trine Grønhaug Halvorsen

Immunocapture LC-MS/MS is a promising technique to ensure high sensitivity and selectivity of low-abundant protein biomarkers. For this purpose, the use of monoclonal antibodies (mAb) is especially attractive as they are renewable reagents that can be standardized. In this article we investigated the possibility of using mAbs developed against intact proteins (anti-protein antibodies) to capture proteotypic epitope peptides. Three mAbs were tested, and all selectively extracted proteotypic epitope peptides from a complex sample. Compared to intact protein extraction, this concept which we call peptide capture by anti-protein antibodies provided cleaner extracts, which further improved the sensitivity. Analysis of three patient samples demonstrated that p can be used for the determination of different endogenous protein levels.

History

Email Address of Submitting Author

trinegha@farmasi.uio.no

Institution

School of Pharmacy, University of Oslo

Country

Norway

ORCID For Submitting Author

0000-0002-2545-3474

Declaration of Conflict of Interest

No conflict of interest

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