A Cocktail of Thermally Stable, Chemically Synthesized Capture Agents for the Efficient Detection of Anti-gp41 Antibodies from Human Sera and Techniques

• Main Author: Pfeilsticker, Jessica A.
• Format: Others
• Published: 2014
• Online Access: https://thesis.library.caltech.edu/8395/61/Thesis.pdf; https://thesis.library.caltech.edu/8395/1/CoverPage.pdf; https://thesis.library.caltech.edu/8395/2/Chapter%201.pdf; https://thesis.library.caltech.edu/8395/3/Chapter%202.pdf; https://thesis.library.caltech.edu/8395/4/Chapter%203.pdf; https://thesis.library.caltech.edu/8395/5/Chapter%204.pdf; https://thesis.library.caltech.edu/8395/28/Appendix%20A.pdf; https://thesis.library.caltech.edu/8395/33/Appendix%20B.pdf; https://thesis.library.caltech.edu/8395/39/Appendix%20C.pdf; https://thesis.library.caltech.edu/8395/49/Appendix%20D.pdf; https://thesis.library.caltech.edu/8395/54/Appendix%20E.pdf; Pfeilsticker, Jessica A. (2014) A Cocktail of Thermally Stable, Chemically Synthesized Capture Agents for the Efficient Detection of Anti-gp41 Antibodies from Human Sera and Techniques. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/0FT2-EF04. https://resolver.caltech.edu/CaltechTHESIS:05262014-170142267 <https://resolver.caltech.edu/CaltechTHESIS:05262014-170142267>

This thesis reports on a method to improve in vitro diagnostic assays that detect immune response, with specific application to HIV-1. The inherent polyclonal diversity of the humoral immune response was addressed by using sequential in situ click chemistry to develop a cocktail of peptide-based capture agents, the components of which were raised against different, representative anti-HIV antibodies that bind to a conserved epitope of the HIV-1 envelope protein gp41. The cocktail was used to detect anti-HIV-1 antibodies from a panel of sera collected from HIV-positive patients, with improved signal-to-noise ratio relative to the gold standard commercial recombinant protein antigen. The capture agents were stable when stored as a powder for two months at temperatures close to 60°C.