Protein Production Shared Resources Core

项目摘要/摘要-蛋白质技术共享资源

PROJECT SUMMARY/ABSTRACT–Protein Technology Shared Resource The goal of the Protein Technology Shared Resource (PTSR) is to generate protein reagents and provide analytical instrumentation that enables SJCCC members to explore the biology and therapeutic consequences of altered protein function in cancer. The PTSR offers state-of-the-art technologies for protein expression and purification from bacteria, insect cells, and mammalian host systems, and provides expert consultation to SJCCC members regarding their needs for these services. The core has recently upgraded its protein expression pipeline by acquiring contemporary automated protein purification systems to enhance the quality of the final product and the documentation of its purity. The PTSR was founded and has been directed by Charles Rock, PhD, an expert in protein expression and purification. Moving forward, the leadership of the PTSR is transitioning in the next funding period to Charalampos Kalodimos, PhD (CBP), the new Chair of the Department of Structural Biology. Kalodimos has decades of experience in protein biochemistry and brings new energy and vision to enhancing the impact of the PTSR on SJCCC research. Currently, Rock and Kalodimos are co- directing the PTSR for one year to ensure a smooth transition in leadership. When new components are fully integrated into the PTSR, the resource will have 9 technical staff, 1 manager, and the Director (Kalodimos). The PTSR is a highly specialized Shared Resource that provides a relatively small but crucial service with a significant impact on the science of the SJCCC. The centralization of resources for protein production, purification, and analysis provides economic value to SJCCC members, and the PTSR can usually offer substantial savings even for commercially available proteins. Cytokines, for example, cost $10 to $30 per microgram commercially, whereas PTSR recently produced 250 micrograms of M-CSF for Kanneganti (CBP) for $0.50 per microgram, saving hundreds of thousands of dollars. Of the 26 manuscripts using the PTSR, 7 (27%) involved intraprogrammatic collaboration, and 4 (15%) involved interprogrammatic collaboration. Work involving the PTSR has had marked scientific impact, including publications from 4 of the 5 Programs, CBP (n=20), DBSTP (n=7), HMP (n=3), and NBTP (n=1). In the upcoming funding period, the PTSR will continue to offer bacterial, insect, and mammalian cell expression systems for protein production, supported by existing fermentation equipment and cell culture facilities. The transition in Directors will be completed, and services will be significantly expanded and improved, including expansion of the protein-expression capabilities and the integration of the molecular-interaction components. State-of-the art cloning and expression services will be augmented by the addition of a Hudson Robotics ProLink Protein Expression System capable of performing high-throughput expression screens of multiple clones, which will be operational by July 2018.