# HYBRIDOMA AND MYELOMA ANTIBODY COMBINING SITES

> **NIH NIH R01** · COLUMBIA UNIV NEW YORK MORNINGSIDE · 1992 · $226,595

## Abstract

The long term goal of this project is to determine the relationship between
diversity and complementarity and to determine the repertoire and fine
specificity of combining sites to a single well defined antigenic
determinant, the site filling epitope of alpha(1 ->6)dextran.  We have
studied a series of hybridomas specific for alpha(1->6)dextran,
characterized their antibodies immunochemically as to fine structure and
idiotypic specificity, and determined the primary structures of their VH
and VL.  Modeling of cavity- and groove-type sites, will be continued to
determine variations in the sites in relation to sequence differences.  An
ultimate objective is to determine the three dimensional xray structure of
their combining sites.  Studies with hybridomas have shown that they do
not comprise the totality of antibody specificities seen in serum.
Thus there is a hidden portion of the repertoire which can only be
discovered by the use of multiple strains, varied conditions of
immunization etc.  This is important in determining the extent of germ line
usage in evaluating the repertoires.  Different VH and VL can generate
similar binding specificities and the presence of N-linked carbohydrate in
VH influences the strength of the antigen-antibody interaction; thus not
only the amino acid sequence of the variable region but also the
carbohydrate present there is important for determining binding.  Site
directed mutagenesis will be coupled with gene transfection techniques to
evaluate the contributions of residues in VH and VL to specificity and
binding.  We have studied a monoclonal anti-blood group A in humans and
mice and find that identity in fine structure can occur with different
residues in the CDRs of the two species; this will be expanded to define
the repertoire in two different species to anti-B as well anti-A in
relation to identities and differences in CDRs and fine specificity of the
sites.  We shall compare the hybridoma repertoire in great detail with that
produced in E. coli by the Lerner procedure with respect to cavity- vs
groove-type sites, binding constants, site sizes, fine structure using
various oligo- and fluorodeoxy oligosaccharides, and idiotype
specificities.
The findings will establish whether the two repertoires are drawn from the
same universe of antibody combining sites.

## Key facts

- **NIH application ID:** 3128472
- **Project number:** 5R01AI019042-08
- **Recipient organization:** COLUMBIA UNIV NEW YORK MORNINGSIDE
- **SAM.gov UEI:** F4N1QNPB95M4
- **Principal Investigator:** ELVIN A KABAT
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1992
- **Award amount:** $226,595
- **Award type:** 5
- **Project period:** 1983-09-30 → 1996-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3128472

## Citation

> US National Institutes of Health, RePORTER application 3128472, HYBRIDOMA AND MYELOMA ANTIBODY COMBINING SITES (5R01AI019042-08). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3128472. Licensed CC0.

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