Abstract

This proposal involves studies by quantitative immunochemical methods of the combining sites of hybridoma antibodies specific for Alpha1 greater than 6 and Alpha1 greater than 3 dextrans, and purification of the Alpha1 greater 6 specific antidextrans in quantities suitable for intensive efforts at crystallization. In addition using recombinant DNA techniques, cDNA clones derived from the H and L chain mRNA of the hybridomas will be sequenced so that specificity differences and X-ray crystallographic data can be correlated with amino acid sequence differences. A set of synthetic Alpha1 greater than 6 glycolipids made by coupling the isomaltose oligosaccharides to stearylamine will be used to produce hybridomas. Their sites will be characterized and studied as above. Rabbit and mouse anti-idiotypic antibodies will be produced to the hybridoma anti-1 yield 6 dextrans and used to define and characterize their idiotypic determinants. Attempts will be made to prepare hybridomas producing monoclonal anti-idiotypic antibodies. Comparison of germ line and expressed genes may make it possible to define the contributions of V, D, J usage, gene conversion and somatic mutation to antibody complementarity and idiotypy.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI019042-03
Application #
3128469
Study Section
(SSS)
Project Start
1983-09-30
Project End
1986-08-31
Budget Start
1985-09-01
Budget End
1986-08-31
Support Year
3
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027

  Publications

Wu, A M; Wu, J H; Chen, Y Y et al. (1999) Further characterization of the combining sites of Bandeiraea (Griffonia) simplicifolia lectin-I, isolectin A(4). Glycobiology 9:1161-70
Wu, J H; Song, S C; Chen, Y Y et al. (1998) Multi-antennary Gal beta1-->4GlcNAc and Gal beta1-->3GalNAc clusters as important ligands for a lectin isolated from the sponge Geodia cydonium. FEBS Lett 427:134-8
Wu, A M; Song, S C; Chang, S C et al. (1997) Further characterization of the binding properties of a GalNAc specific lectin from Codium fragile subspecies tomentosoides. Glycobiology 7:1061-6
Wu, A M; Wu, J H; Song, S C et al. (1996) Bandeiraea (Griffonia) simplicifolia lectin-I, isolectin A4, reacting with Tn (Ga1NAc alpha1 --> Ser/Thr) or galabiose (Ga1 alpha1 --> 4Ga1) containing ligands. FEBS Lett 398:183-6
Nickerson, K G; Tao, M H; Chen, H T et al. (1995) Human and mouse monoclonal antibodies to blood group A substance, which are nearly identical immunochemically, use radically different primary sequences. J Biol Chem 270:12457-65
Wu, A M; Song, S C; Wu, J H et al. (1995) Affinity of Bandeiraea (Griffonia) simplicifolia lectin-I, isolectin B4 for Gal alpha 1-->4 Gal ligand. Biochem Biophys Res Commun 216:814-20
Liao, J; Nickerson, K G; Bystricky, S et al. (1995) Characterization of a human monoclonal immunoglobulin M (IgM) antibody (IgMBEN) specific for Vi capsular polysaccharide of Salmonella typhi. Infect Immun 63:4429-32
Chen, J; Borden, P; Liao, J et al. (1992) Variable region cDNA sequences of three mouse monoclonal anti-idiotypic antibodies specific for anti-alpha(1----6)dextrans with groove- or cavity-type combining sites. Mol Immunol 29:1121-9
Schiffer, M; Kabat, E A; Wu, T T (1992) Subgroups of Tcr alpha chains and correlation with T-cell function. Immunogenetics 35:224-34
Padlan, E A; Kabat, E A (1991) Modeling of antibody combining sites. Methods Enzymol 203:3-21

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