Real-time association rate constant measurement using combination tapered fiber-optic biosensor (CTFOB) dip-probes.
Real-time association rate constant measurement using combination tapered fiber-optic biosensor (CTFOB) dip-probes.
复制标题
使用组合锥形光纤生物传感器 (CTFOB) 浸入式探针进行实时关联速率常数测量。
DOI：
10.1117/12.843266
复制
发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Kapoor,Rakesh
中科院分区：
文献类型：
--
作者：
Simmonds,Boris;Wang,Chun-Wei;Kapoor,Rakesh
本文报道了一种使用基于逝去波的组合锥形光纤生物传感器(CTFOB)DIP探针来测量抗体-抗原相互作用的结合速率常数(Ka)的新方法。通过测定牛血清白蛋白(BSA)的结合速率常数和抗BSA抗体的相互作用证明了该方法的有效性。检测采用“直接法”，将羊抗BSA“捕获”抗体固定在探针表面，用Alexa 488染料直接标记BSA。然后将探针浸入3 NM标记的牛血清白蛋白(1 mg/ml)中。记录的荧光信号与BSA抗BSA偶联物成正比，并使用光纤光谱仪(Ocean Optics，Inc.)获得连续信号。采用476 nm半导体激光器作为激发源。从作为时间函数的信号曲线图中估计缔合常数。室温下测得抗牛血清白蛋白与牛血清白蛋白结合的结合速率常数ka为(8.33±0.01)×104M−1 S−1。
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 3 nM 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) ×104 M−1 s−1.