Hybridoma Facility

杂交瘤设施

基本信息

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
项目类别：
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

项目摘要

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.

杂交瘤设施的目的是生产单克隆抗体（mAb），以用于表征，检测，检测，以及净化肿瘤抗原和其他与癌症相关的蛋白质。该设施主要通过融合与骨髓瘤融合伴侣的抗原免疫小鼠的脾脏，使抗原特异性抗原分泌不朽细胞（杂交瘤）。这种方法的优点是它利用了免疫系统的能力完整的动物，生成与具有高亲和力的抗原反应的抗体，可用于多种需要抗体以高特异性和灵敏度与靶抗原相互作用的应用。研究人员经常为该设施提供高度免疫原性的谷胱甘肽-S-转移酶（GST）融合蛋白，在几乎所有情况下，它都会产生一个有用的分泌IgG的mAB，以识别靶抗原。最多最近的资金期，该设施产生了三类mab：1）识别转录因子和转录因子结构域用于肿瘤发生的细胞过程； 2）针对分子的mab 参与MicroRNA的RNA编辑，这些MicroRNA已成为基因表达的关键调节剂。 3）抗iDotiotypic 用于癌症治疗的mAB。因此，该设施为多个癌症中心提供了独特的试剂具有提高能力进行尖端研究的成员癌症的治疗方法。在最近的资金期间，该设施引入了空心纤维生物反应器为了增强其产生大量mAb的能力，并开始执行定制标签具有荧光色素和其他标签的抗体，以满足研究人员对标有试剂的不断增长的需求用于流式细胞仪和其他分析系统。正在开发的另一个领域是人类mab的产生，使用新方法结合流式细胞术，通过Toll样受体配体激活B细胞，并与永生化爱泼斯坦巴尔病毒。该技术提供了开发可用于治疗人类的试剂的潜力在用小鼠治疗的患者中发展的抗小鼠免疫反应的癌症和旁路问题 mabs。该技术还提供了分析可能在疾病中发展的人类B细胞反应的机会癌症等州。