Preprint Article Version 1 This version is not peer-reviewed

Successive Optimization of a Homogeneous Immunoassay for Antibody Detection in Viral Infections

Version 1 : Received: 21 August 2018 / Approved: 21 August 2018 / Online: 21 August 2018 (05:57:40 CEST)

How to cite: Franco, L.S.; Mujica, V.; Blattman, J.N.; García, A.A. Successive Optimization of a Homogeneous Immunoassay for Antibody Detection in Viral Infections. Preprints 2018, 2018080370 (doi: 10.20944/preprints201808.0370.v1). Franco, L.S.; Mujica, V.; Blattman, J.N.; García, A.A. Successive Optimization of a Homogeneous Immunoassay for Antibody Detection in Viral Infections. Preprints 2018, 2018080370 (doi: 10.20944/preprints201808.0370.v1).

Abstract

Although there are extensive literature reports on the use of gold nanoparticle (AuNP) based homogeneous assays for detection of biomolecules, very few experimental description and procedures involving their preparation are described. In this study, AuNPs conjugated to Bovine Serum Albumin or Envelope protein from Dengue II were developed as a homogeneous immunoassay for antibody detection. We report here optimization of key parameters to prepare an immunoassay like conjugation protein concentration, centrifugation time, electrolyte addition and assay temperature.  We determined that saturating protein concentrations improved AuNPs surface coverage and uniformity of the assay and addition of sodium chloride improved sensitivity of the antibody detection method and assay stability.  Furthermore, we showed that dynamic light scattering can be used to monitor changes in gold nanoparticles in the preparation and detection steps. Additionally, numerical simulations of the plasmonic optical response of AuNPs were carried out to scan for size-dependent response of the AuNPs. The AuNPs homogeneous immunoassay developed was further used in the detection of antibodies in vitro to detect Dengue virus infection.

Subject Areas

gold nanoparticles; homogeneous immunoassay; antibody detection; virus infection; Dengue Protein E

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