A simple approach to improve the sensitivity of enzyme-linked immunosorbent assay: using the IMAPlate 5RC96 for result readout.
In this article, we describe a new enzyme-linked immunosorbent assay (ELISA) setup to improve the sensitivity of commercial or homemade ELISAs. In the new ELISA setup, an IMAPlate 5RC96, a disposable multi-utility lab device developed by NCL New Concept Lab is used as a self-uptaking microcuvette array to read out the result of the ELISA that is performed in the normal 96-well plate with reduced substrate solution and stop solution. A commercial interleukin-6 (IL-6) ELISA reagent kit was used for the evaluation. Compared with the conventional ELISA setup, the new ELISA setup could easily increase the absorbance values by up to more than 10-fold. Therefore, the sensitivity (change in absorbance/change in concentration [DeltaAbs/DeltaConc]) is increased accordingly. In addition, methods to extend the upper detection limit of plate readers for the IMAPlate 5RC96 are described. This new ELISA setup may be more notable for the approach employed than for the specific analyte. It should generally be applicable to any conventional ELISA and should serve as an example of a simple solution that increases the detection sensitivity and/or detection range of other assays as well. We expect the approach to have a substantial practical impact on analytical methods and to accelerate discovery, research, and application of analytes at low concentration in life sciences and diagnostics.
Spies
Peter
P
Institut für Chemie und Bioanalytik, Hochschule für Life Sciences, Fachhochschule Nordwestschweiz, CH-4132 Muttenz, Switzerland.
Mueller
Regina
R
Chen
Guo Jun
GJ
Gygax
Daniel
D
D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D015850:Interleukin-6 / Q000032:analysis*; D057230:Limit of Detection; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity
D015850:Interleukin-6; D011933:Reagent Kits, Diagnostic