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Reference Measurement Procedure Development for C-Reactive Protein in Human Serum
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BibTeX
@MISC{Kilpatrick_referencemeasurement,
author = {Eric L. Kilpatrick and David M. Bunk},
title = {Reference Measurement Procedure Development for C-Reactive Protein in Human Serum},
year = {}
}
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Abstract
This paper describes the development of a reference measurement procedure to quantify human C-reactive protein (CRP) in serum using affinity techniques prior to tryptic digestion and liquid chromatography- tandemmass spectrometry (LC-MS/MS) for the certification of reference materials in clinically relevant ranges. The absence of a suitable internal standard for the CRP measurement, necessary to eliminate potential measurement bias in both the affinity purification and trypsin digestion steps, was addressed using the method of standard addition. The standard addition quantification approach was combined with affinity purification, using an anti-CRP monoclonal antibody conjugated to polystyrene beads, trypsin diges-tion of the purified protein, and LC-MS/MS analysis of CRP tryptic peptides. The effectiveness of intact protein
Keyphrases
reference measurement procedure development c-reactive protein human serum affinity purification human c-reactive protein crp tryptic peptide suitable internal standard lc-ms m analysis tryptic digestion reference measurement procedure liquid chromatography tandemmass spectrometry crp measurement standard addition quantification approach intact protein relevant range anti-crp monoclonal antibody purified protein affinity technique trypsin diges-tion potential measurement bias reference material trypsin digestion step standard addition lc-ms m
