CORE--LYMPHOCYTE/HYBRIDOMA

核心--淋巴细胞/杂交瘤

• 批准号: 6203119
• 负责人: WILLIAM SUTHERLAND
• 金额: --
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-07 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6203119
• 关键词: biomedical equipment resource; biomedical facility; enzyme linked immunosorbent assay; hybridomas; lymphocyte; melanoma; monoclonal antibody

Description: (Applicant's Description) The Lymphocyte Culture/Hybridoma Center (LCC) is a research support facility of the University of Virginia School of Medicine and the Cancer Center. The primary function of the LCC is to make available to researchers the most current technology and expertise for the construction and selection of lymphocyte-myeloma hybridomas for the production of monoclonal antibodies. The LCC also provides expertise in the use of these reagents in basic research programs. Services are customized to the specific requirements of individual investigators to optimize the recovery of appropriate antigen specific monoclonal antibodies. Initial discussions are held with an investigator and his staff in order to understand the experimental system employed in his laboratory and to determine the specific applications for which monoclonal antibodies are required. The staff of the LCC then plan and execute all aspects of hybridoma construction and selection and monoclonal antibody production. LCC staff immunize animals, collect sera and develop ELISA assay strategies appropriate for screening large numbers of hybridoma culture supernatants and for the titration of animal sera. Once an appropriate assay system has been developed and used to demonstrate the successful immunization of the necessary animals, fusions are scheduled on a first come first served basis. The center conducts all aspects of cell fusion, assaying by ELISA for specific antibody positive cultures, culturing, cloning, freezing, and recovery of specific antibody producing clones. Investigators receive, upon request, hybridoma culture supernatants and the individual hybridoma clones. The center also provides other extended services using bulk expansion of clones, large scale in vitro production of monoclonal antibodies using gas permeable bags and hollow fiber bioreactor technology, ascites antibody production, isotyping and subisotyping of monoclonal antibodies, long term cryogenic storage of cells, and the routine analytical and preparative purification of monoclonal and polyclonal antibodies by affinity chromatography on recombinant Protein G columns.

产品描述：淋巴细胞培养/杂交中心（LCC）是弗吉尼亚大学医学院和癌症中心的研究支持机构。 LCC的主要功能是为研究人员提供最新的技术和专业知识，用于构建和选择淋巴细胞-骨髓瘤杂交瘤以生产单克隆抗体。 LCC还提供在基础研究项目中使用这些试剂的专业知识。 服务是根据个体研究者的具体要求定制的，以优化适当抗原特异性单克隆抗体的回收。 与研究人员及其工作人员进行初步讨论，以了解其实验室采用的实验系统，并确定需要单克隆抗体的具体应用。 LCC的工作人员然后计划和执行杂交瘤构建和选择以及单克隆抗体生产的所有方面。 LCC工作人员对动物进行免疫接种，收集血清，并开发适用于筛选大量杂交瘤培养上清液和滴定动物血清的ELISA测定策略。 一旦开发出适当的试验系统并用于证明必要动物的成功免疫，则按照先到先得的原则安排融合。该中心进行细胞融合的所有方面，通过ELISA测定特异性抗体阳性培养物，培养，克隆，冷冻和回收特异性抗体产生克隆。 根据要求，研究者接收杂交瘤培养上清液和单个杂交瘤克隆。 该中心还提供其他扩展服务，包括克隆的批量扩增，使用透气袋和中空纤维生物反应器技术进行单克隆抗体的大规模体外生产，腹水抗体生产，单克隆抗体的同种型和亚同种型，细胞的长期低温储存，以及通过重组蛋白G柱上的亲和色谱法进行单克隆和多克隆抗体的常规分析和制备纯化。