ENGINEERING OF ANTIBODY BINDING SITES TO TUMOR ANTIGENS

杂交瘤方法学在肿瘤学中的应用给了巨大的 推动在癌症治疗中使用新的治疗方法。 基于单克隆抗体的治疗方式的明显潜力是 不幸的是，由于难以获得适当的 单克隆抗体及其商业化使用的成本。 作为 革命性的替代方案，这项建议提供了利用蛋白质 工程化以开发“可编程的”抗体结合位点。 在第一阶段，我们将合成一个模型的VH和VL结构域的基因， 亲和抗体 分离表达的蛋白质将允许 Fv结合位点的重组。 这将被彻底定性 在结构和结合性能方面。 并行计算机 随后的分析将允许合理设计的结合位点， 预定的特异性。 在这个项目的后期阶段，VH和VL基因将被改变， 改造原Fv结合位点。 此外，单个多肽 将通过计算设计从Fv结构衍生的结合位点 并通过重组DNA方法执行。 这个基因 结合位点对于融合到细胞毒性剂的基因是理想的，例如 蓖麻毒素A亚基 我们将培育这样一个嵌合体物种进行试验 体外研究。

The application of hybridoma methodology to oncology has given enormous impetus to the use of novel therapeutic approaches in cancer treatment. The obvious potential of monoclonal-antibody-based treatment modalities is unfortunately limited by difficulties involved with obtaining appropriate monoclonals and with the cost of commercializing their use. As a revolutionary alternative, this proposal offers utilization of protein engineering to develop "programmable" antibody binding sites. In Phase I we will synthesize genes for VH and VL domains of a model high affinity antibody. Isolation of the expressed proteins will permit reassembly of the Fv binding site. This will be thoroughly characterized with respect to structural and binding properties. Concurrent computer analysis will subsequently permit rational design of binding sites of predetermined specificity. In later stages of this project the VH and VL genes will be altered to refashion the original Fv binding site. Furthermore, a single polypeptide binding site derived from the Fv structure will be designed by computation and executed by recombinant DNA methodology. The single gene for this binding site is ideal for fusion to the gene for a cytotoxic agent, such as ricin A subunit. We will generate such a chimeric species for trial studies in vitro.