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A Lectin-Based Gold Nanoparticle Assay for Probing Glycosylation of Glycoproteins

Published

Author(s)

Germarie Sanchez-Pomales, Todd A. Morris, James Falabella, Michael J. Tarlov, Rebecca A. Zangmeister

Abstract

We report a glycoanalysis method in which lectins are used to probe the glycans of glycoproteins that are adsorbed on gold nanoparticles. A model mannose-presenting glycoprotein, ribonuclease B (RNase B), and the therapeutic monoclonal antibody (mAb) Rituxan®, were found to adsorb spontaneously and non-specifically to bare gold nanoparticles such that glycans were accessible for lectin binding. Addition of a multivalent binding lectin, such as concanavalin A (Con A), to a solution of the modified gold nanoparticles resulted in cross-linking of the nanoparticles. This phenomenon was evidenced within 1 minute by a change in the hydrodynamic radius, DH, measured by dynamic light scattering (DLS) and a shift and increase in absorbance of the plasmon resonance band of the gold nanoparticles. By combining the sugar-binding specificity and the cross-linking capabilities of lectins, the non-specific adsorption of glycoproteins to gold surfaces, and the unique optical reporting properties of gold nanoparticles, a glycosylation pattern of RNase B and Rituxan® could be generated. This assay provides advantages over currently used glycoanalysis methods in terms of short analysis time, simplicity of the conjugation method, convenience of simple spectroscopic detection, and feasibility of providing glycan characterization of the protein drug product by using a variety of binding lectins.
Citation
Biotechnology and Bioengineering
Volume
109
Issue
9

Keywords

glycoprotein, glycan, assay, antibody, lectin, gold nanoparticle

Citation

Sanchez-Pomales, G. , Morris, T. , Falabella, J. , Tarlov, M. and Zangmeister, R. (2012), A Lectin-Based Gold Nanoparticle Assay for Probing Glycosylation of Glycoproteins, Biotechnology and Bioengineering, [online], https://doi.org/10.1002/bit.24513 (Accessed May 26, 2024)

Issues

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Created April 17, 2012, Updated November 10, 2018