A comparison of nanoparticle-antibody conjugation strategies in sandwich immunoassays

Justina O. TamWinchester Engineering and Analytical Center, U.S. Food and Drug Administration, Winchester, Massachusetts

Helena de PuigDepartment of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts

Chun-wan YenWinchester Engineering and Analytical Center, U.S. Food and Drug Administration, Winchester, Massachusetts;Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, Massachusetts

Irene BoschInstitute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, Massachusetts

Jose Gómez-MárquezMIT Little Devices Lab and the MIT-SUTD International Design Centre, Massachusetts Institute of Technology, Cambridge, Massachusetts

Charles ClavetWinchester Engineering and Analytical Center, U.S. Food and Drug Administration, Winchester, Massachusetts

Kimberly Hamad-SchifferliDepartment of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts;Department of Engineering, University of Massachusetts Boston, Boston, MassachusettsCorrespondence[email protected]

http://orcid.org/0000-0002-4839-3179

Lee GehrkeInstitute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, Massachusetts;Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MassachusettsCorrespondence[email protected]

http://orcid.org/0000-0002-9387-8212

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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.

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A comparison of nanoparticle-antibody conjugation strategies in sandwich immunoassays

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A comparison of nanoparticle-antibody conjugation strategies in sandwich immunoassays

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