Critical role of the sample matrix in a point-of-care protein chip for sepsis

Journal of Immunological Methods
Ursula SauerClaudia Preininger

Abstract

Both highly specific antibodies and appropriate assay buffers are key elements in the development of sensitive multi-analyte diagnostic tests and essential assay components to minimize interferences from the sample matrix. Herein, we investigate the influence of 0.1 M Tris (pH 7.4)/0.1 M NaCl/10 mM CaCl(2)/0.1% Tween-20 used as assay buffer and diluent for serum, plasma and saliva samples in a protein biomarker chip for the diagnosis of sepsis. In detail, on-chip sandwich assays for detection of IL-6 and PCT are established using pure assay buffer and serum, plasma, and saliva, each diluted by a factor of 10 and 100 with assay buffer. The dilution linearity as well as the cross-reactivity to immobilized IL-8, IL-10 and TNF-α antibodies (<1.8% in plasma and serum) is investigated; furthermore the influence of immunoglobulin G, fibrinogen and lysozyme, highly abundant proteins in serum, plasma and saliva. This effect is two times more pronounced in serum than in plasma and saliva and strongly decreases with increasing analyte concentration. Though the matrix proteins bind unspecifically to the immobilized receptors, they do not prevent the analyte binding; on the contrary, the analyte is reliably detected with high sensitivity, f...Continue Reading

References

Oct 12, 2002·Journal of Medical Virology·Niels FiskerPeer Brehm Christensen
Dec 19, 2002·Molecular & Cellular Proteomics : MCP·N Leigh Anderson, Norman G Anderson
May 22, 2004·Caries Research·A Van Nieuw AmerongenE C I Veerman
Feb 24, 2005·Lab on a Chip·Nicolaos ChristodoulidesJohn T McDevitt
Sep 22, 2005·Lab on a Chip·Chu-Ya YangCarlo D Montemagno
Feb 9, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Evrim Banu Altintaş, Adil Denizli
Aug 26, 2011·Analytical Biochemistry·Ursula SauerClaudia Preininger
Jan 1, 2004·Indian Journal of Clinical Biochemistry : IJCB·Praveen Kumar Shetty, T N Pattabiraman

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Citations

Sep 25, 2012·Expert Opinion on Drug Metabolism & Toxicology·Sarah C CampbellCatherine Mt Sherwin
Apr 12, 2016·Journal of Immunoassay & Immunochemistry·Aleksangr Georgievich PoltavchenkoOleg Viacheslavovich Nechitaylo
Aug 9, 2016·Journal of Virological Methods·Alexander G PoltavchenkoVadim N Gureyev

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