Antigen-antibody interactions: binding studies with fluorescence and surface plasmon resonance exemplified by acid-traseolide as antigen.
The interaction between the musk fragrance acid-traseolide and monoclonal antibodies (mAB) generated against this odorant has been investigated with two different techniques. Fluorescence spectroscopy was used to study the quenching of tryptophan fluorescence of the antibody upon binding acid-traseolide. This spectroscopic approach is based on measurements under equilibrium conditions. The second technique exploited the surface plasmon resonance (SPR) phenomenon. The acid-traseolide was immobilized in the surface matrix and upon presenting mAB changes in SPR were recorded in real time during the association reaction. The SPR approach can be considered as a kinetic method. Although having a different origin, both methods lead to comparable equilibrium dissociation constants (Kd). However, the results obtained with fluorescence spectroscopy were more accurate and reproducible. Not only the association of acid-traseolide with antibody was evaluated, also Fab fragment and peptide (H3-peptide) mimicking the heavy chain CDR3 of this antibody were included in this study. The Kd-values, determined by both methods, increase in the order mAB < Fab < H3-peptide because of diminishing recognition.
Visser
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NV
Unilever Research Laboratorium Vlaardingen, The Netherlands. nina.visser@laser.bc.wau.nl
Smit-Kingma
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IE
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions / Q000276:immunology*; D022801:Complementarity Determining Regions; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D007189:Indans / Q000737:chemistry / Q000276:immunology*; D013050:Spectrometry, Fluorescence / Q000379:methods; D020349:Surface Plasmon Resonance / Q000379:methods
C110695:5-acetyl-1,1,2,6-tetramethyl-3-isopropylindane; D000911:Antibodies, Monoclonal; D022801:Complementarity Determining Regions; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D007189:Indans