The goal of the Antibody Technology Resource Center (ATRC) is to develop technologies to generate high quality recombinant antibodies (rAbs) to high-value protein targets that have proven difficult for Ab generation. Abs are essential reagents for determining how proteins function under normal and pathophysiological conditions yet are not available for much of the proteome and when available are of -variable quality. The generation of widely available, renewable, validated and standardized sets of rAbs will significantly accelerate studies of protein function. We are focusing on generating rAbs to secreted extracellular and membrane proteins and protein post-translational modifications since: 1) they comprise 20-40% of all proteins and 2) they have proven challenging to either express, purify and/or successfully use for Ab generation. The ATRC at the University of California, San Francisco (UCSF) has three Technology Research and Development (TR&D) projects that will develop novel high throughput antigen and antibody generation technology to generate rAbs. Project 1) Develop technology to generate rAbs to secreted extracellular, single pass and multipass membrane proteins using yeast displayed antigens, cell lines and phage antibody libraries, Project Leader: James D. Marks;Project 2) Develop technology to generate functional rAbs to proteases that modulate protease function, Project Leader: Charles Craik;and Project 3) Develop technology to generate rAbs to post-translational modifications and their conformers, Project Leader: James Wells. Driving biomedical projects at UCSF and elsewhere will inform target selection, provide some of the antigens and use and validate rAbs generated by the Center for structural and functional assays. These ongoing NIH-funded projects will benefit from ready access to high quality rAbs. Training in the technologies will be accomplished by hosting an annual symposium, workshops, webinars, and by onsite training of visiting investigators in the use of technology. Dissemination of reagents will be accomplished by publication of methods, and by distribution of plasmids containing antibody genes, antibody vectors, antibody expressing cell lines and antibody reagents.
Antibodies are essential reagents for studying protein function, yet are not available for many proteins and, where available, vary in quality. This significantly limits the study of protein function. Development of technologies to robustly generate renewable and validated antibodies that can be widely distributed will significantly accelerate the rate of biomedical research.
|Pan, Yuchen; Sackmann, Eric K; Wypisniak, Karolina et al. (2016) Determination of equilibrium dissociation constants for recombinant antibodies by high-throughput affinity electrophoresis. Sci Rep 6:39774|
|Wong, Kenneth R; Menendez, Elizabeth; Craik, Charles S et al. (2016) In vivo imaging of protease activity by Probody therapeutic activation. Biochimie 122:62-7|
|Kim, JungMin; Wu, Shenping; Tomasiak, Thomas M et al. (2015) Subnanometre-resolution electron cryomicroscopy structure of a heterodimeric ABC exporter. Nature 517:396-400|
|LeBeau, Aaron M; Sevillano, Natalia; Markham, Kate et al. (2015) Imaging active urokinase plasminogen activator in prostate cancer. Cancer Res 75:1225-35|
|Hornsby, Michael; Paduch, Marcin; Miersch, Shane et al. (2015) A High Through-put Platform for Recombinant Antibodies to Folded Proteins. Mol Cell Proteomics 14:2833-47|
|Li, Keyu; Zettlitz, Kirstin A; Lipianskaya, Julia et al. (2015) A fully human scFv phage display library for rapid antibody fragment reformatting. Protein Eng Des Sel 28:307-16|
|Le Gall, Marianne; CrÃ©pin, Ronan; Neiveyans, Madeline et al. (2015) Neutralization of KIT Oncogenic Signaling in Leukemia with Antibodies Targeting KIT Membrane Proximal Domain 5. Mol Cancer Ther 14:2595-605|