Protein Production Shared Resources Core

项目摘要/摘要-蛋白质技术共享资源 蛋白质技术共享资源(PTSR)的目标是产生蛋白质试剂并提供 分析仪器，使SJCCC成员能够探索生物学和治疗后果 癌症中蛋白质功能改变的研究。PTSR提供了最先进的蛋白质表达和 从细菌、昆虫细胞和哺乳动物宿主系统中提纯，并为SJCCC提供专家咨询 会员对这些服务的需求。核心最近升级了它的蛋白质表达 管道通过收购当代自动化蛋白质纯化系统来提升最终质量 产品及其纯度的文件。PTSR是由查尔斯·洛克创立并指导的， 他是蛋白质表达和纯化方面的专家。展望未来，PTSR的领导层正在过渡 在下一个供资期间，Charalampos Kalodimos博士(CBP)将担任 结构生物学。Kalodimos在蛋白质生物化学方面拥有数十年的经验，并带来了新的能源和 加强PTSR对SJCCC研究的影响的愿景。目前，Rock和Kalodimos正在联合- 指导PTSR一年，以确保领导层的平稳过渡。当新组件完全安装好时 该资源并入临时特别报告，将有9名技术人员、1名管理人员和主任(卡洛迪莫斯)。这个 PTSR是一种高度专业化的共享资源，它提供相对较小但至关重要的服务，具有重要的 对SJCCC科学的影响。蛋白质生产、纯化和生产资源的集中 分析为SJCCC成员提供了经济价值，PTSR通常甚至可以提供大量的节省 以获取商业上可用的蛋白质。例如，细胞因子的商业价格为每微克10至30美元， 尽管PTSR最近以每微克0.50美元的价格为Kannganti(CBP)生产了250微克M-CSF， 节省了数十万美元。在使用PTSR的26篇稿件中，有7篇(27%)涉及 方案内协作，4个(15%)涉及方案间协作。涉及到的工作 PTSR产生了显著的科学影响，包括来自5个方案中的4个的出版物，CBP(n=20)，DBSTP 7例，HMP(3例)，NBTP(1例)。在即将到来的资助期间，PTSR将继续提供细菌、 现有发酵支持的用于蛋白质生产的昆虫和哺乳动物细胞表达系统 设备和细胞培养设施。董事的过渡将完成，服务将显著 扩展和改进，包括扩展蛋白质表达能力和整合 分子相互作用组分。最先进的克隆和表达服务将通过 添加Hudson Robotics ProLink高通量蛋白表达系统 多个克隆的表达屏幕，将于2018年7月投入使用。

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.