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Surface Passivation for Single-molecule Protein Studies

Surface Passivation for Single-molecule Protein Studies

Published on: April 24, 2014

Nanogel surface coatings for improved single-molecule imaging substrates.

Lee A Tessler¹, Casey D Donahoe, Daniel J Garcia

¹Department of Genetics and Center for Genome Sciences, Washington University School of Medicine, 4444 Forest Park Parkway, Campus Box 8510, St. Louis, MO 63108, USA.

Journal of the Royal Society, Interface

|February 18, 2011

Summary

New PEG-BSA nanogel coatings significantly reduce protein and DNA adsorption for sensitive single-molecule bioanalytical applications. This hybrid surface offers superior performance over existing BSA and poly(ethylene glycol) coatings.

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Area of Science:

Biomaterials Science
Surface Chemistry
Analytical Chemistry

Background:

Protein adsorption on surfaces hinders bioanalytical applications.
Bovine serum albumin (BSA) and poly(ethylene glycol) (PEG) coatings minimize non-specific protein adsorption.
PEG-BSA nanogels offer a hybrid approach to low-background surface coatings.

Purpose of the Study:

To characterize PEG-BSA nanogel coatings for single-molecule (SM) protein studies.
To evaluate the nanogel coating's performance against existing BSA and PEG coatings.
To assess the nanogel coating's compatibility with surfactants and its utility in digital immunoassays.

Main Methods:

Single-molecule (SM) adsorption counting to quantify protein adsorption.
Adsorption assays for DNA and assessment of surfactant compatibility.

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Application in a digital immunoassay for single ligand molecule detection.

Main Results:

Nanogel-coated surfaces demonstrated lower protein adsorption than BSA or PEG monolayers.
The nanogel coating exhibited resistance to DNA adsorption.
The nanogel coating was compatible with surfactants, unlike BSA coatings.
High sensitivity and specificity were achieved in detecting single tethered ligand molecules.

Conclusions:

PEG-BSA nanogel coatings provide exceptionally low protein adsorption for SM analysis.
These nanogels are versatile for various SM experiments, including DNA adsorption resistance.
The coating's compatibility and performance in digital immunoassays highlight its practical utility for advanced bioanalytical studies.