Related Experiment Videos
Recombinant antibodies in bioactive peptide design
C Monfardini1, T Kieber-Emmons, J M VonFeldt
1Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia.
The Journal of Biological Chemistry
|March 24, 1995
Summary
Researchers developed novel analogs of granulocyte-macrophage colony-stimulating factor (GM-CSF) interaction sites using recombinant antibody technology. These analogs mimic critical binding regions, offering new strategies for drug design targeting GM-CSF receptors and immune processes.
Area of Science:
- Immunology
- Molecular Biology
- Pharmacology
Background:
- Granulocyte-macrophage colony-stimulating factor (GM-CSF) plays a crucial role in immune and inflammatory responses.
- GM-CSF receptors are key targets for drug development, necessitating a deep understanding of ligand-receptor interactions.
- Developing analogs of interaction sites is a strategy to dissect critical intermolecular interactions.
Purpose of the Study:
- To apply recombinant antibody technology to create analogs of a specific site on GM-CSF.
- To investigate the potential of these analogs as tools for understanding GM-CSF ligand-receptor interactions.
- To explore the feasibility of using recombinant antibody libraries for generating interaction site analogs.
Main Methods:
- Development of polyclonal antisera and monoclonal antibodies with neutralizing activity against human GM-CSF.
- Generation of a recombinant antibody library using spleen cells from mice immunized with anti-GM-CSF antibodies.
- Screening the library with a neutralizing anti-GM-CSF monoclonal antibody (mAb 126.213) to isolate binding clones.
- Sequencing and structural analysis of a selected clone (23.2) to identify mimicry of GM-CSF residues.
- Synthesis and testing of a peptide analog corresponding to a complementarity determining region (CDR).
Main Results:
- A recombinant antibody clone (23.2) was isolated that inhibited the binding of mAb 126.213 to GM-CSF.
- Structural analysis revealed that clone 23.2 potentially mimics specific amino acids on GM-CSF helices.
- A synthetic peptide analog of CDR I bound mAb 126.213, antagonized GM-CSF binding, and blocked bioactivity.
Conclusions:
- Recombinant antibody technology is a feasible approach for developing analogs of critical interaction sites on GM-CSF.
- These analogs can serve as valuable tools for dissecting molecular interactions and informing pharmacologic design.
- The findings support the use of antibody libraries as a source for generating interaction site mimics.