Molecular Tools Core

分子工具核心

• 批准号: 10271747
• 负责人: DALE F MIERKE
• 金额: $ 31.8万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10271747
• 关键词: Affinity; Awareness; Back; Binding; Biochemical; Biomedical Research; Biotechnology; Cell Line; Cells; Centers of Research Excellence; Charge; Cloning; Collaborations; Collection; Communities; Complement; Complex; Consultations; Cost Analysis; Cost Sharing; Development; Elements; Enzymes; Equipment; Escherichia coli; Eukaryotic Cell; Faculty; Feedback; Foundations; Funding; Future; Goals; Grant; HLA-A gene; Hemagglutinin; Heterogeneity; Histone H3; Human; Incubated; Influenza Hemagglutinin; Insecta; Institutes; Laboratories; Lead; Mammalian Cell; Measures; Methods; Modeling; Molecular; Monoclonal Antibodies; Neoplasm Metastasis; New Hampshire; Peptide/MHC Complex; Phase; Pilot Projects; Plant Resins; Play; Procedures; Production; Productivity; Protein Biochemistry; Proteins; Protocols documentation; Reagent; Recombinants; Recovery; Research; Research Personnel; Research Project Grants; Resources; Role; Scheme; Scientist; Sequence Analysis; Services; Specificity; Strategic Planning; Structure; System; T memory cell; Tail; Techniques; Testing; Training; Universities; Work; base; beta-2 Microglobulin; biophysical properties; college; cost; design; design and construction; experience; faculty research; flexibility; improved; instrumentation; large scale production; molecular imaging; mutant; oncolytic virotherapy; operation; pressure; protein expression; protocol development; quality assurance; recruit; scale up; success; tool; vector

The goal of the Molecular Tools Core (MTC) is to provide biomolecular tools to the bioMT faculty to allow them to better achieve their research goals. During the first funding period the MTC assisted over 200 researchers within the greater biomedical community and played a central role in the advancement of the bioMT junior research project leaders (RPL) to externally funded independent investigators. Our success is based on the centralization of the equipment and expertise for providing high-quality purified proteins and other biomolecular tools, dedicated laboratory work by our expert staff, as well as training of users in the utilization of state-of-the- art instrumentation maintained within the core. To complement the expertise of our faculty Director, we recruited a highly experienced Lead Scientist to oversee the operation of the MTC and to provide expert protein biochemistry support and consultation. Her experience in protein production has served her well particularly as a consultant, providing advice from sequence analysis to expression systems to purification protocols to both experienced and novice users. We have established efficient pipelines for protein production in E. coli and mammalian cells and optimized purification schemes. In collaboration with the Molecular Interactions and Imaging Core (MIIC), we have developed protocols for biochemical and biophysical characterization and QA/QC of the final products. Additionally, during phase 1, the MTC created a bioMT Service Center for chargeback costs of supplies, media and reagents related to eukaryotic cell protein production starting October 1, 2020. In the next funding period, we will continue to provide valuable resources to all four RPLs as detailed in Aim 1. One vital feature will be the addition of standard methods for protein production using insect cell lines (Sf9, Hi5). Aim 2 is to enhance the efficient operation of the core, streamlining the day-to-day operations and overall throughput to better serve the research community. We will build on our collection of resources, including strains, vectors, in-house produced enzymes and resins. As part of our phase II strategy, we will expand our throughput capabilities with respect with construct design, cloning, expression screenings to rapidly identify protein targets, reducing the burden of the bioMT investigators on protein production. As the number of users of the MTC has expanded, it is imperative that we invest in the efficiency of the operation of the core and maintain a robust user base as we transition to a cost-share center to guarantee sustainability. Another key element of our phase II strategy is sustainability. As outlined in Aim 3, we will develop cost-recovery models. Faculty will not be charged during phase II, but we will share quarterly usage with projected costs, so they can include costs in grant submissions prior to phase III. Looking towards the future, the MTC will also support collaborative pilot projects designed to nucleate multi-project grants. Overall, the phase II plan for MTC will continue to catalyze the success of RPLs and other bioMT faculty, drawing our faculty together and laying the foundation for the Institute’s long- term transition to COBRE independence.

分子工具核心 (MTC) 的目标是为 bioMT 教师提供生物分子工具，使他们能够 更好地实现他们的研究目标。在第一个资助期内，MTC 资助了 200 多名研究人员 在更大的生物医学界中，并在青年人bioMT的进步中发挥了核心作用 研究项目负责人（RPL）到外部资助的独立研究人员。我们的成功基于 集中提供高质量纯化蛋白质和其他生物分子的设备和专业知识 工具、我们专家人员的专门实验室工作以及对用户使用最新技术的培训 艺术仪器保留在核心内。为了补充我们的教务主任的专业知识，我们招募了 经验丰富的首席科学家负责监督 MTC 的运作并提供专家蛋白质 生物化学支持和咨询。她在蛋白质生产方面的经验对她很有帮助，特别是 顾问，提供从序列分析到表达系统再到纯化方案的建议 有经验的和新手用户。我们建立了高效的大肠杆菌蛋白质生产管道， 哺乳动物细胞和优化的纯化方案。与分子相互作用和 成像核心 (MIIC)，我们开发了生化和生物物理表征以及 QA/QC 协议 最终产品。此外，在第一阶段，MTC 创建了一个用于退款的 bioMT 服务中心 自 2020 年 10 月 1 日起，与真核细胞蛋白生产相关的用品、培养基和试剂的成本。 在下一个资助期内，我们将继续为目标 1 中详述的所有四个 RPL 提供宝贵的资源。 一项重要功能是增加使用昆虫细胞系（Sf9、Hi5）生产蛋白质的标准方法。 目标 2 是增强核心的高效运营，简化日常运营和整体运营 吞吐量，以更好地服务研究界。我们将利用我们收集的资源，包括菌株、 载体、内部生产的酶和树脂。作为我们第二阶段战略的一部分，我们将扩大吞吐量 在构建设计、克隆、表达筛选方面的能力，以快速识别蛋白质靶标， 减轻 BioMT 研究人员在蛋白质生产方面的负担。随着MTC的用户数量不断增加 扩大后，我们必须投资于核心的运营效率并保持强劲的用户 我们过渡到成本分摊中心以保证可持续性的基础。我们第二阶段的另一个关键要素 战略就是可持续性。正如目标 3 中所述，我们将开发成本回收模型。学院不会收取任何费用 在第二阶段，但我们将分享季度使用量和预计成本，因此它们可以包括赠款成本 第三阶段之前提交的材料。展望未来，MTC还将支持合作试点项目 旨在集中多项目赠款。总体而言，MTC二期计划将继续促进成功 RPL 和其他 bioMT 教职人员的组成，将我们的教职人员聚集在一起，为研究所的长期发展奠定了基础 任期过渡到 COBRE 独立性。