# INTERACTIONS OF IMMUNOGLOBULINS

> **NIH NIH R01** · UNIVERSITY OF PENNSYLVANIA · 1987 · $149,706

## Abstract

This study is concerned with two aspects of the immune response.  The first
relates to the affinity limitations inherent in the expression of the
germ-line immunoglobin gene segments and its significance for the affinity
restriction of IgM antibody.  The second aspect is directed to the
specificity and affinity implications of the differences between
immunogenic peptides in the free state i.e. highly flexible, and in the
constrained state in proteins.  Both projects will utilize hybridomas
producing monoclonal antibody to a defined epitope.  There are now
available 60 cell lines producing anti-dansyl
(5-dimethylaminoaphthalene-1-sulfonate) IgM and IgG antibodies whose
binding constants cover a 100,000-fold range.  From these cell lines mRNA
will be isolated and the cDNA prepared by the primer extension method.
Nucleotide sequencing of the variable regions will be carried out by the
Maxam-Gilbert procedure using this cDNA or by the dideoxy method with the
mRNA and the above primer.  For the studies with peptides hybridomas
producing monoclonal antibodies against synthetic peptides whose sequences
are present on the surface of well-defined proteins will be prepared.
Specifically purified antibody will be obtained from ascites and used in
affinity measurements.  The synthetic peptides will be conjugated to
fluorescent probes and fluorescence methodology employed for the
determination of affinity.  Competitive binding experiments will allow the
determination of the binding constants for the peptides present in a
protein.

## Key facts

- **NIH application ID:** 3124565
- **Project number:** 5R01AI009492-27
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **SAM.gov UEI:** GM1XX56LEP58
- **Principal Investigator:** FRED KARUSH
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1987
- **Award amount:** $149,706
- **Award type:** 5
- **Project period:** 1975-04-01 → 1988-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3124565

## Citation

> US National Institutes of Health, RePORTER application 3124565, INTERACTIONS OF IMMUNOGLOBULINS (5R01AI009492-27). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3124565. Licensed CC0.

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