NanoDLSay: a new platform technology for biomolecular detection and analysis using gold nanoparticle probes coupled with dynamic light scattering

摘要

Most analytical techniques that are routinely used in biomedical research for detection and quantification of biomolecules are time-consuming, expensive and labor-intensive, and there is always a need for rapid, affordable and convenient methods. Recently we have developed a new platform technology for biomolecular detection and analysis: NanoDLSay. NanoDLSay employs antibody-coated gold nanoparticles (GNPs) and dynamic light scattering, and correlates the specific increase in particle size after antigen-antibody interaction to the target antigen concentration. We applied this technology to develop an assay for rapid detection of actin, a protein widely used as a loading control in Western Blot analysis. GNPs were coated with two types of polyclonal anti-actin antibodies, and used in the assay to detect two types of actin: p- and bovine skeletal muscle actin in RIPA buffer. The results of our study revealed some complex aspects of actin binding characteristics, which depended on the type of actin reagent and anti-actin antibody used. A surprising finding was a reverse dose-response relationship between the actin concentration and the average particle size in the assay solution, which we attributed to the effect of RIPA buffer. Our results indicate that RIPA may also interfere in other types of nanoparticle-based assays, and that this interference deserves further study.