A label-free nanoparticle aggregation assay for protein complex/aggregate detection and study
Abbreviated Journal Title
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
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.
"A label-free nanoparticle aggregation assay for protein complex/aggregate detection and study" (2010). Faculty Bibliography 2010s. 7008.