Antifouling surface layers for improved signal-to-noise of particle-based immunoassays.

Langmuir : the ACS Journal of Surfaces and Colloids
Annie ChenMatt Trau

Abstract

A 10-fold improvement in the signal-to-noise (S/N) ratio of an optically encoded silica particle-based immunoassay was achieved through incorporating a protein resistant poly(ethylene glycol) (PEG) surface layer and optimizing antibody immobilization conditions. PEG was activated using 2,2,2-trifluoroethanesulfonyl chloride (tresyl) and required a minimum reaction time of 1.5 h. The activated PEG had a reactive half-life of approximately 5 h when stored in acidified dimethyl sulfoxide (DMSO). By increasing the protein incubation time and concentration, a maximum antibody loading on the particle surface of 1.6 x 10(-2) molecules per nm(2) was achieved. The assay S/N ratio was assessed using a multiplexed multicomponent optically encoded species-specific immunoassay. Encoded particles were covalently grafted or nonspecifically coated with either bovine or mouse IgG for the simultaneous detection of complementary anti-IgG "target" or uncomplementary anti-IgG "noise". The versatility and potential as a serum-based assay platform was demonstrated by immobilizing either a polyclonal antibody or an engineered single-chain variable fragment (scFv) capture probe on particles for the detection of the ovarian cancer biomarker, mesothelin ...Continue Reading

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Citations

Jul 21, 2011·Chemical Communications : Chem Comm·Darby KozakMatt Trau
Mar 31, 2016·Lab on a Chip·Balakrishnan RajaRichard Willson
Mar 3, 2015·Biotechnology and Bioengineering·Eric OuelletCharles Haynes
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Feb 16, 2021·Advanced Materials·Eunjung KimSeungjoo Haam
Dec 3, 2010·Langmuir : the ACS Journal of Surfaces and Colloids·Ying XueAlan E Mark

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