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J Immunoassay Immunochem. 2017;38(4):355-377. doi: 10.1080/15321819.2016.1269338. Epub 2016 Dec 16.

A comparison of nanoparticle-antibody conjugation strategies in sandwich immunoassays.

Author information

1
a Winchester Engineering and Analytical Center , U.S. Food and Drug Administration , Winchester , Massachusetts.
2
b Department of Mechanical Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts.
3
c Institute for Medical Engineering and Science, Massachusetts Institute of Technology , Cambridge , Massachusetts.
4
d MIT Little Devices Lab and the MIT-SUTD International Design Centre , Massachusetts Institute of Technology , Cambridge , Massachusetts.
5
e Department of Engineering , University of Massachusetts Boston , Boston , Massachusetts.
6
f Department of Microbiology and Immunobiology , Harvard Medical School , Boston , Massachusetts.

Abstract

Point-of-care (POC) diagnostics such as lateral flow and dipstick immunoassays use gold nanoparticle (NP)-antibody conjugates for visual readout. We investigated the effects of NP conjugation, surface chemistries, and antibody immobilization methods on dipstick performance. We compared orientational, covalent conjugation, electrostatic adsorption, and a commercial conjugation kit for dipstick assays to detect dengue virus NS1 protein. Assay performance depended significantly on their conjugate properties. We also tested arrangements of multiple test lines within strips. Results show that orientational, covalent conjugation with PEG shield could improve NS1 detection. These approaches can be used to optimize immunochromatographic detection for a range of biomarkers.

KEYWORDS:

bioconjugation; dengue; dipstick; lateral flow; nanoparticle; rapid test

PMID:
27982728
PMCID:
PMC6202116
DOI:
10.1080/15321819.2016.1269338
[Indexed for MEDLINE]
Free PMC Article

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