ENGINEERING DOUBLE-VARIABLE REGION ANTIBODIES AGAINST HUMAN IMMUNODEFICIENCY VIRUS
Document Type
Presentation
Start Date
22-10-2010 9:00 AM
End Date
22-10-2010 9:15 AM
Description
Monoclonal antibody therapy has proven successful for treatment of severe diseases such as multiple sclerosis and cancer, as well as serving as powerful labeling reagents for laboratory research. However, therapeutic use of monoclonal antibodies is limited by production costs and development of resistance by its targets. We propose to incorporate the variable regions of two antibodies specific for different epitopes on the same HIV Envelope protein into a single antibody in an attempt to create a bi-specific, tetravalent antibody that retains the functional properties of both parental antibodies. By combining a potent neutralizing antibody against gp120 with another against gp41, an anti-Env specific dual-variable region Ab (DVR-Ab) with potentially greater efficacy than either parent alone may be created. Since the anti-gp41 antibody can effectively kill infected cells when conjugated to a toxin as an immunoconjugate, one aim utilizes a secondary immunotoxin assay to test DVR-Abs ability to kill HIV-infected cells in vitro. Another aim uses an MTS assay (cell viability assay) to test the DVR-Ab's ability to neutralize HIV from infecting susceptible cells. Several DVR-Ab versions have been created using linkers which differ only in the number of repeats of a 5 amino acids sequence -GGGGS. Results show that the DVR-Ab is produced intact, is able to bind both antigens by ELISA, and it kills HIV-infected cells; however, the inner domain still does not function as effectively as anticipated. Further modifications in the linker region and the orientation of the variable regions are underway to enhance the function of the inner antigen-binding site.
Recommended Citation
Craig, Ryan and Pincus, S., "ENGINEERING DOUBLE-VARIABLE REGION ANTIBODIES AGAINST HUMAN IMMUNODEFICIENCY VIRUS" (2010). Dr. Joseph M. Moerschbaecher, III Graduate Research Day. 3.
https://digitalscholar.lsuhsc.edu/grad_rs/2010/presentation1/3
ENGINEERING DOUBLE-VARIABLE REGION ANTIBODIES AGAINST HUMAN IMMUNODEFICIENCY VIRUS
Monoclonal antibody therapy has proven successful for treatment of severe diseases such as multiple sclerosis and cancer, as well as serving as powerful labeling reagents for laboratory research. However, therapeutic use of monoclonal antibodies is limited by production costs and development of resistance by its targets. We propose to incorporate the variable regions of two antibodies specific for different epitopes on the same HIV Envelope protein into a single antibody in an attempt to create a bi-specific, tetravalent antibody that retains the functional properties of both parental antibodies. By combining a potent neutralizing antibody against gp120 with another against gp41, an anti-Env specific dual-variable region Ab (DVR-Ab) with potentially greater efficacy than either parent alone may be created. Since the anti-gp41 antibody can effectively kill infected cells when conjugated to a toxin as an immunoconjugate, one aim utilizes a secondary immunotoxin assay to test DVR-Abs ability to kill HIV-infected cells in vitro. Another aim uses an MTS assay (cell viability assay) to test the DVR-Ab's ability to neutralize HIV from infecting susceptible cells. Several DVR-Ab versions have been created using linkers which differ only in the number of repeats of a 5 amino acids sequence -GGGGS. Results show that the DVR-Ab is produced intact, is able to bind both antigens by ELISA, and it kills HIV-infected cells; however, the inner domain still does not function as effectively as anticipated. Further modifications in the linker region and the orientation of the variable regions are underway to enhance the function of the inner antigen-binding site.