The long-term goal of the TR&D 3 project is to develop high-impact, renewable, and open-source antibody reagents for cellular signaling and biomedical research. Specifically, we plan to focus on signaling events marked by important protein post-translational modifications (PTMs) such as phosphorylation and proteolytic zymogen activation events and their attendant protein conformational changes. While some antibodies do exist for specific phosphorylation sites, these antibodies are often unreliable, rarely renewable, and only available to a fraction of the modified sites. In addition, detecting functional forms of specific proteases has not been routinely possible in cells or in vivo due to the lack of conformationally selective antibodies. Furthermore, the industrialization of high-throughput recombinant antibody (rAb) selection and preparation has been thwarted by the absence of reliable robotic expression platforms. Thus, our immediate goals are to generate high-quality, open-source antibody reagents, by using novel phage display technologies and libraries developed by the UCSF Antibiome Center, to PTMs including: (i) phosphorylation sites, (ii) proteolytic neo-epitopes, and their attendant conformational changes, and (iii) alternative rapid antigen expression technologies such as in vitro transcription and translation (IVTT) that support all three TR&D projects. These goals compliment TR&Ds 1 and 2 by focusing on PTMs within key transmembrane proteins

National Institute of Health (NIH)
National Cancer Institute (NCI)
Biotechnology Resource Grants (P41)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-IMM-N (41))
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of California San Francisco
San Francisco
United States
Zip Code
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; Crépin, 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