Antibody immobilization using heterobifunctional crosslinkers

被引:108
作者
Shriver-Lake, LC
Donner, B
Edelstein, R
Breslin, K
Bhatia, SK
Ligler, FS
机构
[1] Geocenters Inc, Ft Washington, MD 20744 USA
[2] USN, Res Lab, Ctr Biomol Sci & Engn, Washington, DC 20375 USA
关键词
antibody immobilization; protein immobilization; crosslinker; fiber optic biosensor; immunosensor;
D O I
10.1016/S0956-5663(97)00070-5
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Covalent attachment of functional proteins to a solid support is important for biosensors. One method employs thiol-terminal silanes and heterobifunctional crosslinkers such as N-succinimidyl 4-maleimidobutyrate (GMBS) to immobilize proteins through amino groups onto glass, silica, silicon or platinum surfaces. In this report, several heterobifunctional crosslinkers are compared to GMBS for their ability to immobilize active antibodies onto glass cover slips at a high density. Antibodies were immobilized at densities of 74-220 ng/cm(2) with high levels of specific antigen binding. Carbohydrate-reactive crosslinkers were also compared to GMBS using a fiber optic biosensor to detect fluorescently-labeled antigen. At the concentrations tested, the antibodies immobilized with carbohydrate-reactive crosslinkers bound more antigen than GMBS immobilized antibodies as indicated by the fluorescence signal. (C) 1997 Published by Elsevier Science Limited.
引用
收藏
页码:1101 / 1106
页数:6
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