Abstract
Enzyme-linked immunosorbent assays (ELISA) show extensive application in immunoassays, to detect and monitor protein biomarkers in clinical diagnosis. Nevertheless, the time required and its multiple steps limit its application. We take advantage of a polyethyleneimine (PEI) gold nanoparticle (GNP) coated microwell plate to perform blocking-free ELISA, in which no nonspecific protein adsorption appears on the GNP layer. If the PEI-GNP coated microwell plate and immobilization of captured antibodies on the plate are prepared in advance, such as using an ELISA kit, the whole ELISA process can be finished in less than 2 h. Meanwhile, we have ensured that the GNP layer can preserve the precision and good linearity of ELISA without causing negative effects on the plate.
Highlights
Proteins are macromolecules performing a vast array of functions within organisms [1].Some proteins are biomarkers related to different kinds of diseases and detection and monitoring of their concentrations are of great significance for clinical diagnosis [2,3,4]
After successful functionalization of the gold nanoparticle (GNP) layer on the microplate, we investigated whether it has positive effects on the commercial enzyme-linked immunosorbent assay (ELISA)
GNP layer on the microplate, we investigated whether it has positive effects on the commercial ELISA. −1 (R = 0.99) between OD620 and known carcinoembryonic antigen (CEA) concentrations
Summary
Proteins are macromolecules performing a vast array of functions within organisms [1]. Blocking is a step for filling a plate to avoid unspecific adsorption of enzyme-labelled antibodies after primary antibodies adsorb on the plate [13,14]. Albumin from bovine serum (BSA), polyethylene glycol (PEG), and skimmed milk powder are the most widely used blocking agents [15,16,17] Their type, concentration, and incubation time influence blocking efficiency a great deal. With recent advances in nanomaterials and Sensorsscience, 2018, 18, 3537 polymer different strategies based on these are applied to an ELISA to increase sensitivity, blocking efficiency, or realize a blocking-free ELISA [20,21,22]. Commercial microwell plates with an in situ blocking synthesized gold nanoparticle layer couldELISA amplify the Commercial signal and lower the plates detection efficiency, or realize a blocking-free [20,21,22]. We maintain the advantages of ELISA such as its high andprecision good linearity, endow and endow new characteristics, including free-blocking and a reduced new characteristics, including free-blocking and a reduced operating timeoperating (Scheme 1).time (Scheme 1)
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