Hybridoma Facility

杂交瘤设施的目标是生产单克隆抗体（MAb），用于表征，检测， 以及纯化肿瘤抗原和其它癌症相关蛋白。该设施主要通过融合 用骨髓瘤衍生的融合配偶体使抗原特异性抗体分泌永生化的抗原免疫小鼠的脾脏 细胞（杂交瘤）。这种方法的优点是它利用了免疫系统的能力， 一个完整的动物，以产生抗体，与抗原反应具有高亲和力，这可以用于各种 需要抗体以高特异性和灵敏度与靶抗原相互作用的应用。 研究人员经常向该机构提供高免疫原性谷胱甘肽-S-转移酶（GST）融合蛋白， 其在几乎所有情况下产生识别靶抗原的有用的IgG分泌性MAb。最 在最近的资助期间，该基金产生了三类MAb：1）识别转录因子的MAb， 用于研究肿瘤发生的细胞过程的转录因子结构域; 2）抗分子的MAb 参与microRNA的RNA编辑，microRNA已成为基因表达的关键调节因子; 3）抗独特型 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 提高了对潜在机制进行尖端研究的能力， 癌症的治疗方法。在最近的资助期间，该设施引进了中空纤维生物反应器 以提高其生产大量单克隆抗体的能力，并开始进行定制标签， 抗体与荧光染料和其他标签，以满足日益增长的需求，研究人员标记试剂， 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克隆抗体的产生， 使用流式细胞术、通过toll样受体配体的B细胞活化和 爱泼斯坦巴尔病毒。该技术提供了开发可用于治疗人类肿瘤的试剂的潜力。 癌症和旁路问题与抗小鼠免疫反应的发展， 单克隆抗体这项技术还提供了分析人类B细胞反应的机会， 国家如癌症。

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.