Variable region cDNA sequences and antigen binding specificity of mouse monoclonal antibodies to isomaltosyl oligosaccharides coupled to proteins. T-dependent analogues of alpha(1----6)dextran

• Published in: The Journal of immunology (1950) Vol. 142; no. 3; pp. 863 - 870
• Main Authors: Matsuda, T, Kabat, EA
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Am Assoc Immnol 01-02-1989; American Association of Immunologists
• Subjects: Amino Acid Sequence; Animals; Antibodies, immunoglobulins; Antibodies, Monoclonal - analysis; Antibody Specificity; Base Sequence; Binding Sites, Antibody; Biological and medical sciences; Carrier Proteins - immunology; Dextrans - immunology; Disaccharides - immunology; DNA - isolation & purification; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Hybridomas - metabolism; Immunoglobulin Heavy Chains - genetics; Immunoglobulin Heavy Chains - isolation & purification; Immunoglobulin Heavy Chains - metabolism; Immunoglobulin kappa-Chains - genetics; Immunoglobulin kappa-Chains - isolation & purification; Immunoglobulin kappa-Chains - metabolism; Immunoglobulin Variable Region - genetics; Immunoglobulin Variable Region - isolation & purification; Immunoglobulin Variable Region - metabolism; Isomaltose - immunology; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Molecular immunology; Molecular Sequence Data; Monoclonal antibodies; T-Lymphocytes - immunology; Immunoglobulins; Nucleotide sequence; Variable region; Rodentia; light-Peptide chain; Hybridoma; Binding site; Monoclonal antibody; Immunochemistry; Enzyme immunoassay; Structure function relationship; Antigen; Dextran; Vertebrata; Mammalia; heavy-»Peptide chain; Complementary DNA; Mouse; Multigene family; Molecular cloning; ELISA assay; Nucleic acid
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.142.3.863

Four mouse hybridomas specific for alpha(1---6)dextran, 16.4.12E (IgA kappa, C57BL/6), 28.4.10A (IgM kappa, BALB/c), 35.8.2H (IgG1 kappa, BALB/c), and 36.1.2D (IgM kappa, BALB/c) were obtained by immunization with the T-dependent Ag isomaltohexaose or isomaltotriose coupled to keyhole limpet hemocyanin or to BSA. Immunochemical characterization of the hybridoma antibodies showed that 16.4.12E and 36.1.2D had cavity-type combining sites, recognizing the terminal non-reducing end of alpha(1---6)dextran, whereas 28.4.10A and 35.8.2H had groove-type sites, recognizing internal linear segments of the dextran. The V region cDNA of the H and L chains of the antibodies were cloned and sequenced. VH of 16.4.12E and VH of 36.1.2D belonged to the X24 and Q52 germ-line gene families, respectively. The VH and V kappa sequences of 16.4.12E and V kappa sequence of 36.1.2D were highly homologous to those of W3129, the only anti-alpha(1---6)dextran mAb with a cavity-type site thus far sequenced; 16.4.12E differed from W3129 in the D, JH, and J kappa. VH genes of 28.4.10A and 35.8.2H were homologous to those of several anti-alpha(1---6)dextrans with groove-type sites, but belonged to the J558 germ-line gene family, differed from the other J558 anti-alpha(1---6)dextrans, probably representing a different germ-line subfamily. The L chain sequence of 28.4.10A encoded by V kappa-Ars and J kappa 2 was almost identical to other groove-type anti-alpha(1---6)dextrans obtained by immunizing with the T-independent glycolipid Ag, stearyl-isomaltotetraose. Use of T-dependent Ag such as isomaltosyl oligosaccharide-protein conjugates provides an additional parameter for probing the fine structure of antibody combining sites and evaluating the V-gene repertoire of anti-alpha(1---6)dextrans.