OTHER FUNCTIONS SBIR PHASE II, TOPIC 288, "RECOMBINANT ANTIBODIES AS AFFINITY CA

该项目的目标是开发针对75种癌症相关蛋白的重组抗体。在I期研究中，我们已经证明可以从大型酵母展示的人抗体文库中分离多个抗体克隆。分离的抗体表现出与源自杂交瘤技术的单克隆抗体相当的高亲和力（IOW nM至亚nM）和高特异性。同时，优化了技术平台以支持酵母细胞中更稳健的全长IgG展示和一致的抗体表达。 在这个拟议的第二阶段项目中，我们建议使用AvantGn专有的酵母展示抗体库分离针对NCI临床蛋白质组学技术倡议开发的75种癌症相关抗原的抗体。分离的抗体将在亲和力（开和关结合常数）和特异性方面进行表征，并在常用测定如免疫沉淀和/或蛋白质印迹中进行性能表征，目的是鉴定适合于开发ELISA的每个抗原的2个不同抗体克隆。这些抗体将与NCI的临床蛋白质组学技术倡议合作，进一步整合到蛋白质组学研究平台中。

The objective of this proposed project is to develop recombinant antibodies against 75 cancer-related proteins. In the Phase I study, we have demonstrated that multiple antibody clones could be isolated from a large yeast displayed human antibody library. The isolated antibodies exhibit high affinity (lOW nM to sub-nM) and high specificity compariable to monoclonal antibodies derived from hybridoma technology. In parallel, the technology platform was optimzed to support more robust full length IgG display and consistent antibody expression in yeast cells. In this proposed Phase II project, we propose to isolate antibodies against 75 cancer related antigens developed by NCI's Clinical Proteomic Technologies Initiative using AvantGn's proprietary yeast display antibody library. The isolated antibodies will be characterized in terms of affinity (on and off binding constants) and specificity, and for performance in commonly used assays such as immunoprecipitation, and/or Western blot, with the goal of identifying 2 distinct antibody clones per antigen suitable for developing ELiSAs. These antibodies will be further intergrated into proteomic research platforms in collabration with NCI's Clinical Proteomic Technologies Initiative.