Immunoassay reagents for thyroid testing.: 3.: Determination of the solution binding affinities of a T4 monoclonal antibody Fab fragment for a library of thyroxine analogs using surface plasmon resonance
Immunoassay reagents for thyroid testing.: 3.: Determination of the solution binding affinities of a T4 monoclonal antibody Fab fragment for a library of thyroxine analogs using surface plasmon resonance
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DOI：
10.1021/bc9701353
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发表时间：
1998-01-01
影响因子：
4.7
通讯作者：
Pan, Y
中科院分区：
文献类型：
--
作者：
Adamczyk, M;Johnson, DD;Pan, Y
制备了甲状腺素类似物和示踪物文库，并利用BIAcore表面等离子体共振仪检测了它们与抗t -4 Fab片段的溶液结合亲和力。通过测定溶液中已知浓度的游离抗t -4 Fab片段与生物传感器表面的初始结合率，对高密度L-T-4模拟生物传感器进行校准。随后将一系列单独的甲状腺素类似物和示踪剂浓度与固定浓度的抗t -4 Fab片段混合。测定了平衡状态下各溶液中游离抗- t -4 Fab片段的浓度，并推导了每种情况下的平衡解离常数K-D。将溶液中测定的K-D值与在BIAcore仪器上使用单个生物传感器表面进行直接动力学分析所测定的值进行比较。
A library of thyroxine analogs and tracers was prepared, and their solution binding affinities for an anti-T-4 Fab fragment were determined using a single high-density L-T-4 biosensor surface ina BIAcore surface plasmon resonance instrument. The high-density L-T-4 analog biosensor was calibrated by determination of the initial binding rate was of known concentrations of free anti-T-4 Fab fragment in solution to the biosensor surface. A range of individual thyroxine analog and tracer concentrations was subsequently mixed with a fixed concentration of anti-T-4 Fab fragment. The concentration of free anti-T-4 Fab fragment in each solution at equilibrium was determined, and the equilibrium dissociation constant (K-D) for each case was derived. The K-D values determined in solution are compared to values determined by a direct kinetic analysis on the BIAcore instrument using individual biosensor surfaces.