Optimization of fluoroimmunoassay against C-reactive protein exploiting immobilized-antigen glass slide.

An optimization experiment for an indirect-competitive (IC) fluoroimmunoassay (FIA) against C-reactive protein (CRP) was conducted exploiting an immobilized-antigen glass slide and an anti-CRP antibody tagged with fluorescent silica nanoparticles (FSNPs). The optimized conditions for the IC FIA were as follows: time and concentration of treatment with glutaraldehyde, 30 min and 1.5%, respectively; time of reaction with coating antigen and concentration of coating antigen for immobilization, 1 h and 0.1 mg/mL, respectively; concentration of FSNP-anti-CRP antibody conjugate coupled by the biotin-avidin interaction, the bioconjugate, for immune reaction, 0.250 mg/mL; concentration of bovine serum albumin (BSA) for blocking and time of blocking with BSA, 3% and 30 min, respectively. By using the glass slide, a highly sensitive detection against CRP was possible with the limit of detection below 0.1 ng/mL.