Development of Assay Platforms for Monitoring Human C-Reactive Protein

It is imperative to have rapid diagnostic assays available that provide healthcare workers with a rapid yes/no or semi-quantitative response to a biomarker of interest. Agglutination assays use the clustering of antibodies in the presence of an antigen to semi-quantitatively monitor specific protein levels. Our research details the development of a latex agglutination test (LAT) for the detection of C-reactive protein (CRP), an important marker of inflammation in various disease states, at physiologically-relevant levels. Blue latex beads were covalently coated with Protein A for oriented antibody immobilization, then functionalized with anti-CRP antibody at various concentrations. We determined the optimal concentrations of Protein A and anti-CRP and will carry out an assessment of particle size effects on visualization and detectable CRP levels. Further experiments will improve this test’s sensitivity and develop this into a platform assay for other biomarkers. For quantitative measurements in a laboratory environment, we also have developed a sandwich ELISA assay for CRP using anti-CRP conjugated with alkaline phosphatase. These platforms will establish and optimize the immunoassays, then we’ll use novel nanoparticles such as fluorescent liposomes and europium chelate particles to provide greater sensitivity and ease of use versus traditional latex beads or enzymes used for signal amplification.https://orb.binghamton.edu/research_days_posters_spring2020/1020/thumbnail.jp