SBIR Phase II: Antigen-Mediated Selection of Hybridomas

这个小企业创新研究(SBIR)第二阶段项目旨在开发一种快速、灵敏和高度特异的方法，通过结合单细胞凝胶微滴(GMD)封装技术、一种新的蛋白质捕获格式和荧光激活细胞分选来生产单抗和发展杂交瘤细胞系。以胰岛素为模型抗原，第一阶段研究表明，基于分泌抗体的分泌水平和抗原特异性，可以快速分离组成异质群体1%亚群的单个细胞。第二阶段研究将通过允许同时分析其他抗体特性，包括抗体同型和封闭特性，来优化检测格式。利用新融合的杂交瘤，第二阶段的研究将分离和丰富生产克隆，并将结果与传统方法进行比较，这些方法需要耗费时间和劳动强度的限制稀释克隆。单抗被广泛用作研究、治疗、诊断和成像试剂，并越来越多地用于蛋白质组学的新兴领域，用于发现新的药物靶点和定位疾病特异性标记。GMD方法将减少生产时间和成本，提高抗体质量和产量，并允许分离稀有细胞。

This Small Business Innovation Research (SBIR) Phase II project aims to develop a rapid, sensitive and highly specific method for monoclonal antibody production and hybridoma cell line development by combining singlecell gel microdrop (GMD) encapsulation technology, a novel protein capture format, and fluorescence activated cell sorting. Using insulin as a model antigen, Phase I studies demonstrated that individual cells,which comprised a 1% sub-population of a heterogeneous population, could be rapidly isolated based on both secretion level and antigen specificity of the secreted antibody. Phase II research will optimize the assay format by permitting simultaneous analysis of other antibody properties, including antibody isotype and blocking properties. Using newly fused hybridomas, Phase II research will isolate and enrich productiveclones and compare results with conventional methods which require use of time consuming and labor intensive limiting dilution cloning.Monoclonal antibodies are widely used as research, therapeutic, diagnostic, and imaging reagents, and are increasingly used in the emerging field of proteomics for discovering new drug targets and locating disease specific markers. The GMD method will reduce production time and costs, improve antibody quality and yield, and permit isolation of rare cells.