Holographic molecular binding assays

We demonstrate that holographic particle characterization can directly detect binding of proteins to functionalized colloidal probe particles by monitoring the associated change in the particles’ size. This label-free molecular binding assay uses in-line holographic video microscopy to measure the d...

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Bibliographic Details
Published in:Scientific reports 2020-02, Vol.10 (1), p.1932-1932, Article 1932
Main Authors: Zagzag, Yvonne, Soddu, M. Francesca, Hollingsworth, Andrew D., Grier, David G.
Format: Article
Language:English
Subjects:
631/61/32
639/624/1107
Humanities and Social Sciences
Immunoglobulin G
Immunoglobulins
Measurement techniques
Microfluidics
Microscopy
multidisciplinary
Physics
Population
Proteins
Science
Science (multidisciplinary)
Spheres
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Summary:We demonstrate that holographic particle characterization can directly detect binding of proteins to functionalized colloidal probe particles by monitoring the associated change in the particles’ size. This label-free molecular binding assay uses in-line holographic video microscopy to measure the diameter and refractive index of individual probe spheres as they flow down a microfluidic channel. Pooling measurements on 10 4 particles yields the population-average diameter with an uncertainty smaller than 0.5 nm, which is sufficient to detect sub-monolayer coverage by bound proteins. We demonstrate this method by monitoring binding of NeutrAvidin to biotinylated spheres and binding of immunoglobulin G to spheres functionalized with protein A.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-020-58833-7