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Real-time association rate constant measurement using combination tapered fiber-optic biosensor (CTFOB) dip-probes

机译：使用组合锥形光纤生物传感器（CTFOB）浸入探针进行实时关联速率常数测量

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摘要

This document reports a novel method of measuring association rate constant (k_a) 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. Measuredrnassociation rate constant k_a for the binding of BSA with anti-BSA at room temperature is (8.33 + 0.01) × 10~4 M~(-1)s~(-1).
机译：该文献报道了一种新的方法，该方法使用基于e逝波的组合锥形光纤生物传感器（CTFOB）浸入探针来测量抗体-抗原相互作用的缔合速率常数（k_a）。通过测量牛血清白蛋白（BSA）的缔合速率常数和抗BSA抗体相互作用证明了该方法。使用“直接法”进行检测；山羊抗BSA“捕获”抗体固定在探针表面，而抗原（BSA）直接用Alexa 488染料标记。随后将探针浸入卵清蛋白（1 mg / ml）中的3nM标记的BSA中。记录的荧光信号与BSA抗-BSA共轭物成比例，并使用光纤光谱仪（Ocean Optics，Inc.）获取连续信号。 476 nm二极管激光器用作激发源。关联常数是根据信号随时间变化的曲线估算的。在室温下，BSA与抗BSA结合的结合速率常数k_a为（8.33 + 0.01）×10〜4 M〜（-1）s〜（-1）。

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