Optical fiber immunosensor for the real-time analysis of ligand-receptor binding kinetics

Hua Lu; Jianmin Ma; Yujie Zhao; Zu-Hong Lu

doi:10.1117/12.364378

17 September 1999 Optical fiber immunosensor for the real-time analysis of ligand-receptor binding kinetics

Hua Lu, Jianmin Ma, Yujie Zhao, Zu-Hong Lu

Proceedings Volume 3863, 1999 International Conference on Biomedical Optics; (1999) https://doi.org/10.1117/12.364378
Event: International Symposium on Biomedical Optics, 1999, Wuhan, China

Abstract

Real-time immunological interaction analysis is of great importance for the understanding of relationship between the structure and function of immunomolecules. A fluorimetric evanescent wave optical fiber immunosensor has been developed for the quantitative analysis of ligand-reactor binding kinetics in real-time. The specific immunological reaction between IgG molecules and fluorescein isothiocyanate (FITC)-labeled anti-IgG antibodies was chosen as a model system. The optical fiber immunosensor probe used in this study consisted of a piece of silanized quartz optical fiber core on which IgG molecules were immobilized. The immunological reactions were monitored fluorimetrically in situ to follow the reaction dynamics on the optical fiber surface. A gradually increase in fluorescence signal was found upon binding of fluorescein isothiocyanate (FITC)-labeled anti-IgG antibodies to the surface immobilized IgG molecules. This signal was directly related to the surface concentration of the analyte and immunological reaction kinetic properties. Thus any change in analytical signal in this study would reflect the real process of ligand-receptor binding on the optical fiber probe surface. A flow injection system was introduced into the experiments to shift the immunological reaction from mass transport to reaction rate limited. The quantitative information obtained from the initial reaction phase of a immunological reaction was then studied with a theoretical model using one to one binding, which was a significant of the real-time binding events to evaluate the binding kinetic parameters. The association and dissociation rate constants for IgG/anti-IgG antibody were calculated to be 1.4 X 10⁶ M^-1s^-1 and 2.1 X 10^-4s^-1, respectively. The results presented in this paper confirmed that the optical fiber immunosensor described in this study might be used as a tool for rapid and sensitive biospecific interaction analysis.

Citation Download Citation

Hua Lu, Jianmin Ma, Yujie Zhao, and Zu-Hong Lu "Optical fiber immunosensor for the real-time analysis of ligand-receptor binding kinetics", Proc. SPIE 3863, 1999 International Conference on Biomedical Optics, (17 September 1999); https://doi.org/10.1117/12.364378

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