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.
Functional Materials Research Group, Korea Food Research Institute, Songnam 463-746, Republic of Korea. k9130sen@hanmail.net
Namsoo
N
Kim
Yong-Jin
YJ
Cho
D000818:Animals; D000941:Antigens / Q000276:immunology*; D002097:C-Reactive Protein / Q000032:analysis* / Q000276:immunology; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods*; D018796:Immunoconjugates; D051381:Rats
D000941:Antigens; D018796:Immunoconjugates; D002097:C-Reactive Protein