Choi Sue Hyung, Lee Jung Wook, Sim Sang Jun
Department of Chemical Engineering, Sungkyunkwan University, 300 Chunchun-dong, Chanan-Gu, Suwon 440-746, South Korea.
Biosens Bioelectron. 2005 Aug 15;21(2):378-83. doi: 10.1016/j.bios.2004.07.037. Epub 2004 Nov 26.
To enhance the feasibility of surface plasmon resonance (SPR) immunosensor as a tool for diagnosing type I diabetes, we enhanced the sensitivity of immunoresponse for detecting the monoclonal anti-glutamic acid decarboxylase (GAD) antibody by modification of mixed self-assembled monolayers (SAMs). The effects of the different mixed SAMs were evaluated with respect to the degree of streptavidin immobilization, the degree of biotin-GAD immobilization, and the immunoresponse sensitivity. Consequently, the sensitivity of the immunoresponse for the detection of anti-GAD antibody was enhanced as a result of the reduction in steric hindrance brought about by using SAMs of heterogeneous lengths. The immunoresponse for detecting the monoclonal anti-GAD antibody was also enhanced with the reduction of the excess immobilization of biotin-GAD and the minimization of non-specific binding that resulted from the simple substitution of the spacer from a carboxylic-terminated SAM for the hydroxyl-terminated SAM.
为提高表面等离子体共振(SPR)免疫传感器作为诊断I型糖尿病工具的可行性,我们通过混合自组装单分子层(SAMs)修饰来提高检测抗谷氨酸脱羧酶(GAD)单克隆抗体的免疫反应灵敏度。针对链霉亲和素固定程度、生物素-GAD固定程度和免疫反应灵敏度评估了不同混合SAMs的效果。结果,由于使用不同长度的SAMs减少了空间位阻,检测抗GAD抗体的免疫反应灵敏度得以提高。通过减少生物素-GAD的过量固定以及将间隔臂从羧基末端SAM简单替换为羟基末端SAM所导致的非特异性结合最小化,检测抗GAD单克隆抗体的免疫反应也得到了增强。