Sensitive detection of C-reactive protein in serum by immunoprecipitation-microchip capillary gel electrophoresis.

Abstract

Sepsis represents a significant cause of mortality in intensive care units. Early diagnosis of sepsis is essential to increase the survival rate of patients. Among others, C-reactive protein (CRP) is commonly used as a sepsis marker. In this work we introduce immune precipitation combined with microchip capillary gel electrophoresis (IP-MCGE) for the detection and quantification of CRP in serum samples. First high-abundance proteins (HSA, IgG) are removed from serum samples using affinity spin cartridges, and then the remaining proteins are labeled with a fluorescence dye and incubated with an anti-CRP antibody, and the antigen/antibody complex is precipitated with protein G-coated magnetic beads. After precipitation the complex is eluted from the beads and loaded onto the MCGE system. CRP could be reliably detected and quantified, with a detection limit of 25 ng/μl in serum samples and 126 pg/μl in matrix-free samples. The overall sensitivity (LOQ = 75 ng/μl, R(2) = 0.9668) of the method is lower than that of some specially developed methods (e.g., immune radiometric assay) but is comparable to those of clinically accepted ELISA methods. The straightforward sample preparation (not prone to mistakes), reduced sample and reagent volumes (including the antibodies), and high throughput (10 samples/3 h) are advantages and therefore IP-MCGE bears potential for point-of-care diagnosis.

• Authors: Ela Herwig, Martina Marchetti-Deschmann, Christian Wenz, Andreas Rüfer, Heinz Redl, Soheyl Bahrami, Günter Allmaier
• Journal: Analytical biochemistry (Anal Biochem) Vol. 478 Pg. 102-6 (Jun 01 2015) ISSN: 1096-0309 [Electronic] United States
• PMID: 25778394 (Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015 Elsevier Inc. All rights reserved.

Chemical References

• C-Reactive Protein

Topics

• C-Reactive Protein (analysis)
• Electrophoresis, Microchip (methods)
• Humans
• Immunoprecipitation (methods)
• Limit of Detection
• Point-of-Care Systems
• Sepsis (blood)
• Serum (chemistry)