Title

A Label-Free Nanoparticle Aggregation Assay For Protein Complex/Aggregate Detection And Study

Keywords

Dynamic light scattering; GAPDH; Gold nanoparticles; Protein aggregates

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 10-25μg/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. © 2010 Elsevier Inc.

Publication Date

1-1-2010

Publication Title

Analytical Biochemistry

Volume

405

Issue

1

Number of Pages

96-102

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1016/j.ab.2010.06.008

Socpus ID

77955173438 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/77955173438

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