Immunosensor with a controlled orientation of antibodies by using NeutrAvidin-protein A complex at immunoaffinity layer
Immunosensor with a controlled orientation of antibodies by using NeutrAvidin-protein A complex at immunoaffinity layer
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DOI：
10.1016/j.jbiotec.2006.05.010
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发表时间：
2006-11-10
影响因子：
4.1
通讯作者：
Pyun, J. C.
中科院分区：
文献类型：
--
作者：
Chung, J. W.;Park, J. M.;Pyun, J. C.
利用中性亲和素-蛋白A复合物在SPR生物传感器的金表面控制抗体的取向。通过将受体抗体处理到SPR生物传感器的抗生物素蛋白层、NeutrAvidin、蛋白A、NeutrAvidin-Protein A复合物和裸金表面来比较受体抗体（抗hIgG）的表面密度。将配体抗体（hIgG）注射到每个IA层中，并估计每单位受体的配体抗体的结合率作为取向控制的参数。在SPR生物传感器的金表面上的NeutrAvidin-Protein A复合物显示出最高的受体抗体的表面密度以及配体抗体与受体抗体的结合比。还在生物素标记的SAM上制备了NeutrAvidin-蛋白A复合物，并且发现与通过将受体抗体化学偶联至SAM层制备的IA层相比，配体/受体的结合比率显著提高。NeutrAvidin-蛋白A复合物显示出最高的配体抗体结合效率，用于检测称为CEA的癌症标志物。采用NeutrAvidin-Protein A复合物和夹心法进行信号放大，灵敏度提高到裸金表面的1.5倍，检测CEA的最低检测限为30 ng/ml。(c)2006 Elsevier B. V.保留所有权利。
The orientation of antibody was controlled by using NeutrAvidin-protein A complex on the gold surface of SPR biosensor. The surface density of receptor antibody (anti-hIgG) was compared by treatment of receptor antibody to the layer of avidin, NeutrAvidin, protein A, NeutrAvidin-protein A complex and bare gold surface of SPR biosensor. The ligand antibody (hIgG) was injected to each IA layer and the binding ratio of ligand antibody per unit receptor was estimated as a parameter of orientation control. The NeutrAvidin-protein A complex on gold surface of SPR biosensor showed the highest surface density of receptor antibody as well as the binding ratio of ligand antibody per receptor antibody. The NeutrAvidin-protein A complex was also prepared on biotin-labelled SAM, and the binding ratio of ligand per receptor was found to be significantly improved in comparison to the IA layer prepared by chemical coupling of receptor antibody to the SAM layer. The NeutrAvidin-protein A complex which showed the highest efficiency for the binding of ligand antibodies, was applied for the detection of a cancer marker called CEA. By using NeutrAvidin-protein A complex and sandwich assay for signal amplification, sensitivity was improved to be 1.5-fold higher than bare gold surface and the detection of CEA with the detection limit of 30 ng/ml was achieved. (c) 2006 Elsevier B.V. All rights reserved.