24 February 2010 Real-time association rate constant measurement using combination tapered fiber-optic biosensor (CTFOB) dip-probes
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Abstract
This document reports a novel method of measuring association rate constant (ka) for antibody-antigen interaction using evanescent wave-based combination tapered fiber-optic biosensor (CTFOB) dip-probes. The method was demonstrated by measuring association rate constant for bovine serum albumin (BSA) and anti-BSA antibody interaction. "Direct method" was used for detection; goat anti-BSA "capture" antibodies were immobilized on the probe surfaces while the antigen (BSA) was directly labeled with Alexa 488 dye. The probes were subsequently submerged in 3nM Labeled BSA in egg albumin (1 mg/ml). The fluorescence signal recorded was proportional to BSA anti-BSA conjugates and continuous signal was acquired suing a fiber optic spectrometer (Ocean Optics, Inc.). A 476 nm diode laser was use as an excitation source. Association constant was estimated from a plot of signal as a function of time. Measured association rate constant ka for the binding of BSA with anti-BSA at room temperature is (8.33 ± 0.01) x 104 M-1s-1.
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Boris Simmonds, Boris Simmonds, Chun-Wei Wang, Chun-Wei Wang, Rakesh Kapoor, Rakesh Kapoor, } "Real-time association rate constant measurement using combination tapered fiber-optic biosensor (CTFOB) dip-probes", Proc. SPIE 7559, Optical Fibers and Sensors for Medical Diagnostics and Treatment Applications X, 75590Q (24 February 2010); doi: 10.1117/12.843266; https://doi.org/10.1117/12.843266
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