The Effect of Peptide Adsorption on Signal Linearity and a Simple Approach to Improve Reliability of Quantification

Overview

Journal J Proteomics

Publisher Elsevier

Specialty Biochemistry

Date 2013 May 14

PMID 23665148

Citations 5

Authors

Stacey Warwood

Adam Byron

Martin J Humphries

David Knight

Affiliations

Soon will be listed here.

Abstract

Biological Significance: Accurate quantification is central to many aspects of science, especially those examining dynamic processes or comparing molecular stoichiometries. In biological research, the quantification of proteins is an important yet challenging objective. Large-scale quantification of proteins using MS often depends on the comparison of peptide intensities with only a single-level calibrant (as in stable isotope labelling and absolute quantification approaches) or no calibrants at all (as in label-free approaches). For these approaches to be reliable, it is essential that the relationship between signal intensity and concentration is linear, without a significant intercept. Here, we show that peptide adsorption can severely affect this relationship, even under controlled conditions, and we demonstrate simple methodologies that can be used to moderate and predict this effect. These findings thus enable the quantification of proteins with increased robustness and reliability.

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