title: One-step kinetics-based immunoassay for the highly sensitive detection of C-reactive protein in less than 30 min.
abstract: This article reveals a rapid sandwich enzyme-linked immunosorbent assay (ELISA) for the highly sensitive detection of human C-reactive protein (CRP) in less than 30 min. It employs a one-step kinetics-based highly simplified and cost-effective sandwich ELISA procedure with minimal process steps. The procedure involves the formation of a sandwich immune complex on capture anti-human CRP antibody-bound Dynabeads in 15 min, followed by two magnet-assisted washings and one enzymatic reaction. The developed sandwich ELISA detects CRP in the dynamic range of 0.3 to 81 ng ml(-1) with a limit of detection of 0.4 ng ml(-1) and an analytical sensitivity of 0.7 ng ml(-1). It detects CRP spiked in diluted human whole blood and serum with high analytical precision, as confirmed by conventional sandwich ELISA. Moreover, the results of the developed ELISA for the determination of CRP in the ethylenediaminetetraacetic acid plasma samples of patients are in good agreement with those obtained by the conventional ELISA. The developed immunoassay has immense potential for the development of rapid and cost-effective in vitro diagnostic kits.
authors.affiliation: Laboratory for MEMS Applications, Department of Microsystems Engineering-IMTEK, University of Freiburg, 79110 Freiburg, Germany; HSG-Institut für Mikro- und Informationstechnik (IMIT), 79110 Freiburg, Germany. Electronic address: sandeep.kumar.vashist@hsg-imit.de.
authors.forename: Sandeep Kumar
authors.initials: SK
authors.lastname: Vashist
authors.affiliation: HSG-Institut für Mikro- und Informationstechnik (IMIT), 79110 Freiburg, Germany.
authors.forename: Gregor
authors.initials: G
authors.lastname: Czilwik
authors.affiliation: HSG-Institut für Mikro- und Informationstechnik (IMIT), 79110 Freiburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: van Oordt
authors.affiliation: Laboratory for MEMS Applications, Department of Microsystems Engineering-IMTEK, University of Freiburg, 79110 Freiburg, Germany; HSG-Institut für Mikro- und Informationstechnik (IMIT), 79110 Freiburg, Germany.
authors.forename: Felix
authors.initials: F
authors.lastname: von Stetten
authors.affiliation: Laboratory for MEMS Applications, Department of Microsystems Engineering-IMTEK, University of Freiburg, 79110 Freiburg, Germany; HSG-Institut für Mikro- und Informationstechnik (IMIT), 79110 Freiburg, Germany.
authors.forename: Roland
authors.initials: R
authors.lastname: Zengerle
authors.affiliation: Sektion Experimentelle Anaesthesiologie, University Hospital Ulm, 89081 Ulm, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Marion Schneider
authors.affiliation: Innovative Chromatography Group, Irish Separation Science Cluster (ISSC), Department of Chemistry, Analytical and Biological Chemistry Research Facility (ABCRF), University College Cork, Cork, Ireland.
authors.forename: John H T
authors.initials: JH
authors.lastname: Luong
mesh_terms: D002097:C-Reactive Protein / Q000032:analysis*; D003362:Cost-Benefit Analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000191:economics / Q000379:methods*; D006801:Humans; D007700:Kinetics
chemical_list: D002097:C-Reactive Protein
keywords: C-reactive protein; Dynabeads; In vitro diagnostics; Magnet; One-step kinetics; Sandwich ELISA