Hybridoma Facility

杂交瘤设施

• 批准号: 7945021
• 负责人: ANDREW J CATON
• 金额: $ 4.03万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-15 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7945021
• 关键词: Adopted; Affinity; Animals; Antibodies; Antibody Formation; Antigen Targeting; Antigens; Area; Ascites; B-Cell Activation; B-Lymphocytes; Basic Cancer Research; Biochemical; Biological Assay; Bioreactors; Bypass; Cancer Center; Cancer Research Project; Cell Line; Cell Separation; Cell physiology; Cells; Chimeric Proteins; Cloning; Collaborations; Column Chromatography; Complex; Custom; Disease; Enzyme-Linked Immunosorbent Assay; Ethics; Fiber; Flow Cytometry; Fluorochrome; Freezing; Funding; Gene Expression; Gene Expression Regulation; Generations; Glutathione S-Transferase; Goals; Hemorrhage; Human; Human Herpesvirus 4; Hybridomas; Immune response; Immune system; Immunization; Immunoglobulin G; Immunoglobulins; Immunohistochemistry; Immunoprecipitation; In Vitro; Institution; Label; Laboratories; Lead; Life; Ligands; Light; Link; Liquid substance; Malignant Neoplasms; Methods; MicroRNAs; Monitor; Monoclonal Antibodies; Multiple Myeloma; Mus; Nitrogen; Patients; Peptides; Procedures; Production; Proteins; Protocols documentation; Quality Control; RNA Editing; Reagent; Recombinants; Regulator Genes; Research; Research Personnel; Screening procedure; Self Tolerance; Sensitivity and Specificity; Services; Signal Transduction; Specificity; Spleen; Systems Analysis; Techniques; Technology; Therapeutic; Tissue Stains; Toll-like receptors; Tumor Antigens; anticancer research; cancer therapy; cell growth; immunogenic; immunogenicity; meetings; member; novel; repository; response; success; tissue culture; transcription factor; tumorigenesis

The goal of the Hybridoma Facility is to produce monoclonal antibodies (MAbs) for use in characterizing, detecting, and purifying tumor antigens and other cancer-related proteins. The Facility generates MAbs primarily by fusing the spleens of antigen-immunized mice with a myeloma-derived fusion partner to immortalize antigen-specific antibodysecreting cells (hybridomas). The advantage of this approach is that it exploits the ability of the immune system of an intact animal to generate antibodies that react with antigens with high affinity, which can be used in a variety of applications that require the antibody to interact with target antigens with high specificity and sensitivity. Investigators often provide the facility with highly immunogenic glutathione-S-transferase (GST) fusion proteins, which in almost all cases generate a useful IgG-secreting MAb that recognizes the target antigen. In the most recent funding period, the Facility produced three classes of MAbs: 1)MAbs that recognize transcription factors and transcription factor domains for use in studies of cellular processes of oncogenesis; 2) MAbs against molecules involved in RNA editing by microRNAs, which have emerged as crucial regulators of gene expression; and 3) antiidiotypic MAbs for use in cancer therapy. The facility has thus provided unique reagents to multiple Cancer Center members that have advanced their abilities to perform cutting edge research into underlying mechanisms of, and therapeutic approaches to cancer. In the most recent funding period the facility introduced hollow fiber bioreactors to enhance its capacity to produce large quantities of MAbs, and is beginning to perform custom-labeling of antibodies with fluorochromes and other tags to meet the increasing needs of investigators for labeled reagents for use in flow cytometry and other analysis systems. Another area being developed is the generation of human MAbs, using new methods combining flow cytometry, B cell activation via toll-like receptor ligands and immortalization with Epstein Barr virus. This technology provides the potential to develop reagents that can be used to treat human cancers and bypass problems with anti-mouse immune responses that develop in patients treated with mouse MAbs. This technology also offers the opportunity to analyze human B cell responses that may develop in disease states such as cancer.

杂交瘤设备的目标是生产用于鉴定、检测、 以及提纯肿瘤抗原和其他癌症相关蛋白。该设施主要通过融合 与骨髓瘤融合伙伴抗原免疫的小鼠脾永生化抗原特异性抗体分泌 细胞(杂交瘤)。这种方法的优点是它利用了免疫系统的能力 一种完整的动物，能够产生与高亲和力的抗原反应的抗体，这种抗体可以用于各种 要求抗体与靶抗原高度特异性和敏感性相互作用的应用。 研究人员经常向该设施提供高免疫原性的谷胱甘肽-S-转移酶融合蛋白， 在几乎所有的情况下，它都会产生一种有用的分泌免疫球蛋白的单抗，可以识别目标抗原。在大多数情况下 在最近的供资期间，该基金生产了三类单抗：1)识别转录因子的单抗和 转录因子结构域，用于研究肿瘤发生的细胞过程；2)抗分子单抗 参与由microRNAs进行的RNA编辑，microRNAs已成为基因表达的关键调节因素；以及3)抗独特型 用于癌症治疗的单抗。该设施因此为多个癌症中心提供了独特的试剂 提高了对潜在机制进行前沿研究的能力的成员，以及 癌症的治疗方法。在最近的筹资期间，该设施引入了中空纤维生物反应器。 以提高其大量生产单抗的能力，并开始对其进行定制标签 带有荧光染料和其他标记的抗体，以满足研究人员对标记试剂日益增长的需求 用于流式细胞仪和其他分析系统。另一个正在开发的领域是人类单抗的产生， 使用新的方法结合流式细胞术，通过Toll样受体配体激活B细胞和永生化 爱泼斯坦-巴尔病毒。这项技术为开发可用于治疗人类的试剂提供了潜力 用小鼠治疗的患者出现的抗鼠免疫反应的癌症和旁路问题 单抗。这项技术还提供了分析人类B细胞反应的机会，这些反应可能会在疾病中发展 癌症等州。