Automated System for High-Throughput In Vitro Selection of Recombinant Antibodies

用于重组抗体高通量体外选择的自动化系统

• 批准号: 8247377
• 负责人: JAMES A WELLS
• 金额: $ 60万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-15 至 2014-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8247377
• 关键词: Antibodies; Antigens; Automation; Bacteriophages; Biological; Biological Assay; Biological Markers; Cells; Communities; Diversity Library; Equipment; Eukaryotic Cell; Funding; Future; Generations; Genes; Human; Image; Immunohistochemistry; Immunoprecipitation; In Vitro; Minor; Pattern; Policies; Positioning Attribute; Proteins; Proteome; Reagent; Recombinant Antibody; Research; Specificity; System; Technology; Therapeutic; Tissues; United States National Institutes of Health; Western Blotting; Yeasts; high throughput screening; in vivo; instrument; instrumentation; programs; protein function; small molecule; technology development

DESCRIPTION (provided by applicant): We are requesting funds to purchase high throughput automation for the in vitro selection and characterization of recombinant antibodies. The antibodies will be displayed on phage or yeast, and selected against antigens that are either purified or displayed on eukaryotic cells. These antibodies will serve as biological probes, biomarkers and potential therapeutics. There is currently no appropriate instrumentation for this technology at UCSF or nearby campuses. The addition of this high-throughput automation would therefore serve a unique purpose at UCSF, while supporting at least five NIH-funded projects. The new equipment would be placed in the Small Molecule Discovery Center (SMDC) at UCSF and be available to the entire UC community. The SMDC has already established the space, staff, and recharge policies to maintain this equipment. The SMDC includes a high-throughput screening (HTS) group and is therefore well-positioned to maximize the use of new automation for technology development. Antibodies are essential reagents for determining how proteins function under normal or pathophysiological conditions. Uses of antibodies include quantifying proteins, identifying the temporal and spatial pattern of expression in cells and tissue, and identifying interacting partners. Such studies require antibodies of high specificity that function in assays including Western blotting, immunoprecipitation, immunohistochemistry (IHC) and in vivo imaging. Over half the human proteome is not annotated and functional antibodies are not reliably available for these proteins. Many of the limitations of traditional antibody generation can be overcome by the use of display technologies and antibody (Ab) gene diversity libraries. The purchase of this instrument has broad support from three major users with well-established research programs from three departments at two different UCSF campuses. The automation would support the NIH funded research of these users and future minor users throughout UCSF.

描述（由申请人提供）：我们正在申请资金购买高通量自动化设备，用于重组抗体的体外选择和表征。抗体将展示在噬菌体或酵母上，并针对纯化或展示在真核细胞上的抗原进行选择。这些抗体将用作生物探针、生物标志物和潜在的治疗方法。目前加州大学旧金山分校或附近校园没有适合该技术的仪器。因此，增加这种高通量自动化将为 UCSF 提供独特的用途，同时支持至少五个 NIH 资助的项目。新设备将放置在加州大学旧金山分校的小分子发现中心（SMDC），并可供整个加州大学社区使用。 SMDC 已经制定了维护该设备的空间、人员和充电政策。 SMDC 包括一个高通量筛选 (HTS) 小组，因此能够最大限度地利用新的自动化技术开发。抗体是确定蛋白质在正常或病理生理条件下如何发挥作用的重要试剂。抗体的用途包括定量蛋白质、识别细胞和组织中表达的时间和空间模式以及识别相互作用的伙伴。此类研究需要具有高特异性的抗体，可在蛋白质印迹、免疫沉淀、免疫组织化学 (IHC) 和体内成像等检测中发挥作用。超过一半的人类蛋白质组没有注释，并且这些蛋白质的功能性抗体无法可靠地获得。传统抗体生成的许多限制可以通过使用展示技术和抗体 (Ab) 基因多样性库来克服。该仪器的购买得到了三个主要用户的广泛支持，这些用户拥有来自加州大学旧金山分校两个不同校区的三个部门的成熟研究项目。自动化将支持 NIH 资助的这些用户和整个 UCSF 未来未成年用户的研究。

项目成果

A High Through-put Platform for Recombinant Antibodies to Folded Proteins.

• DOI: 10.1074/mcp.o115.052209
• 发表时间: 2015-10
• 期刊: Molecular & cellular proteomics : MCP
• 作者: Hornsby M;Paduch M;Miersch S;Sääf A;Matsuguchi T;Lee B;Wypisniak K;Doak A;King D;Usatyuk S;Perry K;Lu V;Thomas W;Luke J;Goodman J;Hoey RJ;Lai D;Griffin C;Li Z;Vizeacoumar FJ;Dong D;Campbell E;Anderson S;Zhong N;Gräslund S;Koide S;Moffat J;Sidhu S;Kossiakoff A;Wells J
• 通讯作者: Wells J