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

Analytical Biochemistry
Ela HerwigGünter Allmaier

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...Continue Reading

References

Apr 18, 2001·Analytical Chemistry·L BousseR Dubrow
Dec 19, 2002·Molecular & Cellular Proteomics : MCP·N Leigh Anderson, Norman G Anderson
Sep 23, 2003·Clinical Chemistry·Natalie KhuseyinovaWolfgang Koenig
Aug 13, 2004·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Liliana SimonJacques Lacroix
Jun 18, 2005·Memórias do Instituto Oswaldo Cruz·Fernando A BozzaHugo C Castro Faria Neto
Aug 9, 2005·Journal of Proteome Research·Jon M JacobsRichard D Smith
Jun 13, 2006·Biosensors & Bioelectronics·Martin H F MeyerMichael Keusgen
Jun 5, 2007·The American Journal of Emergency Medicine·H Bryant Nguyen, Dustin Smith
Aug 12, 2009·Analytical Biochemistry·Oliver PoetzThomas O Joos
Dec 29, 2009·Frontiers in Bioscience (Elite Edition)·Rudolf Anderson, Rene Schmidt
Jan 13, 2010·Clinical Microbiology Reviews·Nicasio ManciniMassimo Clementi
Feb 11, 2010·Critical Care : the Official Journal of the Critical Care Forum·Charalampos Pierrakos, Jean-Louis Vincent
Jun 1, 2010·International Journal of Medical Microbiology : IJMM·Michael Bauer, Konrad Reinhart
Jan 18, 2011·Clinica Chimica Acta; International Journal of Clinical Chemistry·Dan WenRu-Tai Hui

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Citations

Oct 5, 2016·Analytical Chemistry·Brendon SealeAaron R Wheeler
Nov 3, 2016·Journal of Separation Science·Sille Štěpánová, Václav Kašička
May 5, 2017·The Analyst·Mohamed DawodRobert T Kennedy
Nov 3, 2020·Journal of Separation Science·Shannon L ThomasKatarína Maráková

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