Title

A label-free nanoparticle aggregation assay for protein complex/aggregate detection and study

Authors

Authors

J. Bogdanovic; J. Colon; C. Baker;Q. Huo

Comments

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Abbreviated Journal Title

Anal. Biochem.

Keywords

Gold nanoparticles; Dynamic light scattering; Protein aggregates; GAPDH; DYNAMIC LIGHT-SCATTERING; III PNEUMOCOCCUS POLYSACCHARIDE; HOMOLOGOUS; ANTIBODY; PRECIPITIN REACTION; 2-HYBRID SYSTEM; FLUORESCENCE; ASSOCIATION; PROBES; Biochemical Research Methods; Biochemistry & Molecular Biology; Chemistry, Analytical

Abstract

The detection, analysis, and understanding of protein complexes/aggregates and their formation process are extremely important for biomolecular research, diagnosis, and biopharmaceutical development. Unfortunately, techniques that can be used conveniently for protein complex/aggregate detection and analysis are very limited. Using gold nanoparticle immunoprobes coupled with dynamic light scattering (DLS), we developed a label-free nanoparticle aggregation immunoassay (NanoDLSay) for protein aggregate detection and study. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), a protein target used routinely in Western blot as a loading control, is demonstrated here as an example. Through this study, we discovered that GAPDH has a strong tendency to form large aggregates in certain buffer solutions at a concentration range of 10-25 mu g/ml. The strong light scattering property of gold nanoparticles immunoprobes greatly enhanced the sensitivity of the dynamic light scattering for protein complex/aggregate detection. In contrast to fluorescence techniques for protein complex and aggregation study, the protein targets do not need to be labeled with fluorescent probe molecules in NanoDLSay. NanoDLSay is a very convenient and sensitive tool for protein complex/aggregate detection and study. (C) 2010 Elsevier Inc. All rights reserved.

Journal Title

Analytical Biochemistry

Volume

405

Issue/Number

1

Publication Date

1-1-2010

Document Type

Article

Language

English

First Page

96

Last Page

102

WOS Identifier

WOS:000280890600012

ISSN

0003-2697

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