Antibody & Phage Display Shared Resource

点击翻译按钮获取中文摘要

ANTIBODY & PHAGE DISPLAY SHARED RESOURCE - PROJECT SUMMARY/ABSTRACT The Antibody & Phage Display Shared Resource offers a variety of cutting-edge technologies to develop antibodies for use as key research reagents. The Shared Resource is led by Faculty Head Adrian Krainer, Ph.D, and Director Johannes Yeh, Ph.D. The Shared Resource collaborates with Cancer Center members to address fundamental and compelling questions in cancer biology, in research areas including: (1) The understanding of the tumor microenvironment; (2) The discovery of surface biomarkers and their differential expression during tumorigenesis; and (3) Receptor signaling in cancer. Shared Resource staff work closely with scientists to select the best approach for their experimental needs, and specialized screens are developed in close coodination with individual scientists. The Shared Resource also tests and implements new techniques, reagents, and assay improvements related to the different stages of antibody production. Technologies currently available in the Shared Resource include hybridoma development by fusion of rodent spleen cells with immortalized cell lines, generation of nanobodies from immunized alpacas, and direct cloning of the VH-VL domains from B cells from immunized rodents or alpacas into expression plasmids. Yeast or phage display techniques are used to optimize the binding affinities of antibodies identified after the first round of immunization and selection. A library of hybridomas generated in the Shared Resource is maintained, and staff will produce and purify antibodies from these lines on request. In the last funding period this Shared Resouce was used by twelve Cancer Center members (32%), and has contributed towards seven publications by members. Novel antibodies have generated reagents that have been used for genomics (e.g., ChIP), cell separation (e.g., FACS), and single-cell analysis. Finally, recently developed antibodies are currently being investigated for potential diagnostic or therapeutic applications by members, and are thus playing an essential role in cancer programs at CSHL.