Characterization of monoclonal antibodies against glucosyltransferase synthesizing water-insoluble glucan from Streptococcus sobrinus B13.
Four hybrid cell lines secreting monoclonal antibodies (McAbs) against glucosyltransferase synthesizing water-insoluble glucan (GTase-I) were generated by fusion of myeloma cells (P3U1) with splenocytes from mice immunized with GTase-I from Streptococcus sobrinus B13-N. Cell lines 29E7, 21GC7, and 42HB8 were found to secrete an IgG2a-type immunoglobulin, and 29EG, an IgM-type immunoglobulin. These two isotypes of McAbs were used for the determination of the binding sites on the GTase molecule, with use of an enzyme-linked immunosorbent assay. The binding site for McAb 29EG was different from the site that bound other McAbs. McAb 29EG was found to inhibit water-insoluble glucan synthesis by GTase-I by 50%, and 29E7 was found to inhibit it weakly. However, other McAbs did not show any inhibitory effect in spite of binding to GTase-I. McAb 42HB8 strongly inhibited GTase-I-mediated adherence of heat-killed cells of S. sobrinus B-13N to glass surfaces. When the McAbs were tested for their cross-reactivity among GTase preparations from different mutans streptococci, McAb 29EG reacted with S. cricetus and S. sobrinus, but not with other mutans streptococci. Three other McAbs, 21GC7, 29E7, and 42HB8, were found to react only with the enzyme from S. sobrinus. These findings indicated that the specificity of the McAbs studied varied with respect to the antigenic sites on the GTase-I molecule, and that some of the sites differed in their functions.
Ochiai
K
K
Department of Microbiology, Nihon University, School of Dentistry at Matsudo, Chiba, Japan.
Fukushima
K
K
Shiota
T
T
Ikeda
T
T
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D001422:Bacterial Adhesion; D001666:Binding Sites, Antibody / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D005936:Glucans / Q000096:biosynthesis; D005964:Glucosyltransferases / Q000037:antagonists & inhibitors / Q000276:immunology* / Q000378:metabolism; D012703:Serotyping; D013291:Streptococcus / Q000201:enzymology / Q000276:immunology; D013295:Streptococcus mutans / Q000201:enzymology / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005936:Glucans; C036655:1,3-alpha-D-glucan synthase; D005964:Glucosyltransferases