title: Analysis of the interaction between an alpha (1----6)dextran-specific mouse hybridoma antibody and dextran B512 by affinity electrophoresis.
abstract: Carbohydrates are common environmental antigens. As dextran B512 is composed of a repeating structure of simple antigenic determinants, it is widely used to study the immunochemical properties of immunoglobulins. Two-dimensional affinity electrophoresis patterns of a mouse monoclonal antidextran antibody (35.8.2H; IgG1, BALB/c) were produced to obtain insights into the microheterogeneity of the monoclonal antibody. The monoclonal antibody was separated into about six spots which had an identical affinity to dextran B512, but differed in their isoelectric points (pI). In addition, the pH dependence of the binding affinity of this antidextran to dextran B512 was examined. By comparing affinities obtained by affinity electrophoresis between weakly basic (pH 9.5) and weakly acidic (pH 3.8) discontinuous buffer systems, the latter showed an affinity about 500 times lower than the former. The change in the affinity was investigated with a continuous pH gradient by an affinity titration curve and was seen to change markedly at about pH 6. This suggests that the histidine at residue 34 in the light-chain CDR1 is largely responsible for the dextran binding.
authors.affiliation: First Department of Biochemistry, Yamaguchi University School of Medicine, Ube, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Mimura
authors.forename: K
authors.initials: K
authors.lastname: Nakamura
authors.forename: K
authors.initials: K
authors.lastname: Takeo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology; D002846:Chromatography, Affinity; D003911:Dextrans; D004591:Electrophoresis, Polyacrylamide Gel; D006825:Hybridomas / Q000276:immunology*; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D051379:Mice; D008807:Mice, Inbred BALB C; D008815:Mice, Inbred Strains
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D003911:Dextrans