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）是促进生物医学发展的基本工具。 研究和理解，检测和治疗癌症。Fox Chase癌症中心（FCCC）杂交瘤设施拥有定制杂交瘤生成以及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，能够有效和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，该中心的八个实验室成为该设施的新用户。此外，将细胞储存在该设施维持的液氮冷冻机中的实验室数目从23个增加到26个，从而减少了单个实验室重复使用这些冷冻机的情况。目前，免疫生物学项目以外的研究人员构成了该中心的主要用户群。总体而言，26名研究人员与同行评审的资金（自1999年以来增加了25%）参与了11个研究项目中的8个，代表了该中心的所有三个部门，使用该设施。2003年，经同行审查的该基金使用率为98%。自1999年以来，新的设施服务包括体外生产高滴度mAb（上次审查的目标），通过ELISA定量体外鼠mAb生产，通过玻片电泳监测兔、大鼠和仓鼠mAb的体外生产，mAb纯化，聚丙烯酰胺凝胶电泳以确定纯化mAb的纯度，用荧光染料标记mAb和生产重组IL-6。自上次审查以来，对较老服务的需求也有所增长，包括杂交瘤克隆（增长32%）和mAb同种分型（增长30%）。所提供的服务接近设施工作人员的能力（1.5 FTE）。该设施的未来目标是提高生产兔杂交瘤的效率和成本效益，因为兔通常产生高亲和力抗体，并且是许多研究人员提高抗体的首选。该设施还将产生和选择一组大鼠和兔单克隆抗体，这些抗体识别通常被工程化到重组蛋白中的表位标签。当蛋白质特异性mAb尚不可用时，这种标签特异性mAb可用于检测和纯化标记蛋白质，并且也可用于ELISA。