PROBING BIOSENSOR INTERFACES BY MULTIFREQUENCY PHASE AND MODULATION TOTAL INTERNAL-REFLECTION FLUORESCENCE (MPM-TIRF)

被引:14
作者
BRIGHT, FV
机构
[1] Department of Chemistry, Acheson Hall, State University of New York at Buffalo, Buffalo
关键词
FLUORESCENCE; TIME-RESOLVED FLUORESCENCE OF INTERFACIAL SPECIES; BIORECEPTORS; AFFINITY CONSTANTS; BIOSENSORS;
D O I
10.1366/0003702934067919
中图分类号
TH7 [仪器、仪表];
学科分类号
0804 ; 080401 ; 081102 ;
摘要
The multifrequency phase and modulation (MPM) technique is used in concert with total internal reflection fluorescence (TIRF) to probe in situ the distribution of free and analyte-bound sites of a fluorescently labeled, silica-immobilized F(ab') antibody fragment. Using this new technique, one is able to recover the affinity constant (K(f)) for the immobilized antibody, determine the effects of storage time on K(f) and the excited-state fluorescence decay parameters, and follow time-dependent changes in antibody conformation at the silica substrata. This new technique adds a powerful tool to the study of biosensor interfaces.
引用
收藏
页码:1152 / 1160
页数:9
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