Fiber optic biosensor for the detection of C-reactive protein and the study of protein binding kinetics.
Application of a fiber optic biosensor (FOB) to the real-time investigation of the interaction kinetics between FITC-conjugated monoclonal sheep anti-human C-reactive protein (CRP) antibody and CRP isoforms on the surface of optical fiber is described. Recently, both the native pentameric CRP (pCRP), an acute phase protein belonging to pentraxin family, and an isoform of pCRP, modified CRP (mCRP), have been suggested to have proinflammation effects on vascular cells in acute myocardial infarction (AMI). In current studies, we generate mCRP from pCRP, and use several methods including fluorescence spectral properties, circular dichroism, analytical ultracentrifuge, and Western blotting to demonstrate their differences in physical and chemical properties as well as the purity of pCRP and mCRP. In addition, we design and implement an FOB to study the real-time qualitative and quantitative biomolecular recognition of CRP isoforms. Specifically, the association and dissociation rate constants of the reaction between FITC-conjugated monoclonal sheep anti-human CRP antibody and the pCRP and mCRP are determined. The feasibility of our current approach to measure the association and dissociation rate constants of the reaction between tested CRP isoforms was successfully demonstrated.
Chou
Chien
C
National Yang-Ming University, Institute of Biophotonic Engineering, and Institute of Radiological Sciences, Taipei 112, Taiwan. cchou@ym.edu.tw
Hsu
Hsien-Yeh
HY
Wu
Hsieh-Ting
HT
Tseng
Kai-Yu
KY
Chiou
Arthur
A
Yu
Chih-Jen
CJ
Lee
Zheng-Yuan
ZY
Chan
Tsu-Shin
TS
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002097:C-Reactive Protein / Q000032:analysis* / Q000276:immunology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005336:Fiber Optic Technology / Q000295:instrumentation* / Q000379:methods; D007118:Immunoassay / Q000295:instrumentation*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D055100:Optical Fibers; D011485:Protein Binding; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D002097:C-Reactive Protein