CORE--HYBRIDOMA

核心--杂交瘤

• 批准号: 7142516
• 负责人: Robert C Young
• 金额: $ 6.1万
• 依托单位: FOX CHASE CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-10 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7142516
• 关键词: biomedical facility; hybridomas; immunologic substance development /preparation; monoclonal antibody; neoplasm /cancer; neoplasm /cancer immunology

Monoclonal antibodies (mAb) are fundamental tools for the advancement of biomedical research and the understanding, detection and treatment of cancer. The Fox Chase Cancer Center (FCCC) Hybridoma Facility has the expertise and resources both for the custom generation of hybridomas and for the production and purification of mAb. The Facility enables a diverse group of scientists, especially those unfamiliar with the specialized methods required to produce hybridomas, to obtain valuable antibody reagents efficiently and economically. As a reflection of this assistance, eight laboratories in the Center have become new users of the Facility since 1999. In addition, the number of laboratories storing cells in the liquid nitrogen freezers maintained by the Facility expanded from 23 to 26, thus reducing the duplication of these freezers in individual laboratories. Currently, investigators outside the Immunobiology Program comprise the major user group in the Center. Overall, 26 investigators with peer-reviewed funding (25% increase since 1999) participating in 8 of the 11 Research Programs, representing all three Divisions of the Center, use the Facility. In 2003, peer-reviewed usage of the Facility was 98%. Since 1999, new Facility services include in vitro production of high-titer mAb (a goal of the last review), quantitation of in vitro murine mAb production by ELISA and monitoring of in vitro production of rabbit, rat and hamster mAb by slide-electrophoresis, purification of mAb, polyacrylamide gel electrophoresis to determine the purity of purified mAb, labeling mAb with fluorescent dyes and production of recombinant IL-6. Demand since the last review has also grown for older services including hybricloma cloning (32% increase) and mAb isotyping (30% increase). Services provided are at near capacity for the Facility staff (1.5 FTE). A future goal of the Facility is to improve the efficiency and cost-effectiveness of producing rabbit hybridomas, as rabbits usually produce high affinity antibodies and are the first choice among many investigators for raising antibodies. The Facility will also generate and select a panel of rat and rabbit monoclonal antibodies that recognize epitope tags that are often engineered into recombinant proteins. Such tag-specific mAb could be used for detection and purification of tagged proteins when a protein-specific mAb is not yet available and would also be useful for sandwich-ELISA.

单克隆抗体 (mAb) 是推动生物医学进步的基本工具 研究以及癌症的理解、检测和治疗。 Fox Chase 癌症中心 (FCCC) 杂交瘤设施拥有用于杂交瘤定制生成以及 mAb 生产和纯化的专业知识和资源。该设施使不同的科学家群体，特别是那些不熟悉生产杂交瘤所需的专门方法的科学家，能够高效、经济地获得有价值的抗体试剂。作为这种援助的反映，自 1999 年以来，该中心的 8 个实验室已成为该设施的新用户。此外，该设施维护的液氮冷冻柜中储存细胞的实验室数量从 23 个增加到 26 个，从而减少了重复这些冰箱在各个实验室中。目前，免疫生物学项目之外的研究人员构成了该中心的主要用户群体。总体而言，有 26 名获得同行评审资金的研究人员（自 1999 年以来增加了 25%）参与 11 个研究项目中的 8 个项目，代表该中心的所有三个部门，使用该设施。 2003 年，经过同行评审的该设施的使用率为 98%。自 1999 年以来，新设施服务包括高滴度 mAb 的体外生产（上次审查的目标）、通过 ELISA 对体外鼠 mAb 生产进行定量以及通过玻片电泳监测兔、大鼠和仓鼠 mAb 的体外生产、mAb 的纯化、聚丙烯酰胺凝胶电泳以确定纯化的 mAb 的纯度、用荧光染料标记 mAb 以及重组 IL-6 的生产。自上次审查以来，对旧服务的需求也有所增长，包括杂交瘤克隆（增加 32%）和单克隆抗体同型分型（增加 30%）。所提供的服务已接近设施工作人员的能力（1.5 FTE）。该设施未来的目标是提高兔杂交瘤的生产效率和成本效益，因为兔通常产生高亲和力抗体，是许多研究人员产生抗体的首选。该设施还将生成并选择一组大鼠和兔单克隆抗体，这些抗体可识别通常被设计成重组蛋白的表位标签。当蛋白质特异性 mAb 尚未可用时，此类标签特异性 mAb 可用于检测和纯化标记蛋白，并且也可用于夹心 ELISA。