Molecular recognition of avidin on biotin-functionalized gold surfaces detected by FT-IRRAS and use of metal carbonyl probes.
Fourier transform infrared reflection-absorption spectroscopy (FT-IRRAS) was successively used to monitor the covalent immobilization of biotin molecules onto a planar gold substrate covered with a self-assembled monolayer of cystamine and to transduce the molecular recognition of avidin and biotin. This detection was greatly facilitated and made selective by the labeling of avidin and of biotin with various transition metal carbonyl probes. The binding of avidin to the surface was optimized by blocking the nonspecific binding sites by adsorption of an unrelated protein, bovine serum albumin. This work exemplifies the feasibility of detecting biomolecular associations involving molecules of any size at a liquid/solid interface by using a simple and accessible surface analysis technique.
Yam
Chi-Ming
CM
Laboratoire de Physico-Chimie des Surfaces, CNRS, ESA 7045, Ecole Nationale Supérieure de Chimie de Paris, 11 rue Pierre et Marie Curie, Paris cedex 5, F-75231, France.
Pradier
Claire-Marie
CM
Salmain
Michèle
M
Fischer-Durand
Nathalie
N
Jaouen
Gérard
G
D001360:Avidin / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D015394:Molecular Structure; D009942:Organometallic Compounds / Q000737:chemistry*; D011485:Protein Binding; D012680:Sensitivity and Specificity; D017550:Spectroscopy, Fourier Transform Infrared; D013499:Surface Properties
D004800:Enzymes, Immobilized; D009942:Organometallic Compounds; D001360:Avidin; D001710:Biotin; D006046:Gold