Antigen-antibody binding in reverse micelles: interaction of monoclonal antibodies with a myelin basic protein peptide.
Reverse micelles can be used to mimic biological processes occurring at interfaces. To investigate antigen-antibody binding in a membrane-like environment, we first obtained Fab fragments from monoclonal antibodies against bovine myelin basic protein (MBP), an encephalitogenic protein. The binding of the fragments to a dansylated synthetic human MBP peptide gly(119)-gly(131), presenting sequence homologies with a viral protein, was measured in buffer and for the first time in reverse micelles of sodium bis(2-ethylhexyl) sulfosuccinate, in isooctane. Analysis of the fluorescence polarisation titration curves discloses that the Fab fragments in reverse micelles have retained the high affinity for the peptide found in buffer, and similar to that for intact MBP.
School of Biological and Molecular Sciences, Oxford Polytechnic, Headington, United Kingdom.
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Groome
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Vacher
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Nicot
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Waks
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000276:immunology*; D003102:Colloids; D003619:Dansyl Compounds / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D005454:Fluorescence Polarization; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D008823:Micelles; D008969:Molecular Sequence Data; D008970:Molecular Weight; D004676:Myelin Basic Protein / Q000276:immunology* / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology*; D011994:Recombinant Proteins
D000911:Antibodies, Monoclonal; D000941:Antigens; D003102:Colloids; D003619:Dansyl Compounds; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D008823:Micelles; D004676:Myelin Basic Protein; D010446:Peptide Fragments; D011994:Recombinant Proteins; C065358:dansyl myelin basic protein, Gly(119)-Gly(131)-, synthetic