ENGINEERING OF ANTIBODY BINDING SITES TO TUMOR ANTIGENS

肿瘤抗原抗体结合位点的工程设计

• 批准号: 3491062
• 负责人: JAMES S. HUSTON
• 金额: $ 5万
• 依托单位: CREATIVE BIOMOLECULES, INC.
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-05-01 至 1985-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3491062
• 关键词: X ray crystallography; antibody specificity; antitumor antibody; biotechnology; chemical binding; computer data analysis; conformation; digoxin; genetic manipulation; high performance liquid chromatography; immunoconjugates; immunoglobulin genes; immunoglobulin structure; molecular cloning; monoclonal antibody; neoplasm /cancer immunotherapy; nucleic acid chemical synthesis; nucleic acid sequence; tumor antigens

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.

杂交瘤方法在肿瘤学中的应用给予了巨大的帮助 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。 作为一个 革命性的替代方案，该提案提供了蛋白质的利用 工程开发“可编程”抗体结合位点。 在第一阶段，我们将合成模型高 VH 和 VL 结构域的基因 亲和抗体。 表达蛋白质的分离将允许 Fv 结合位点的重组。 这将被彻底表征 关于结构和结合特性。 并发电脑 随后的分析将允许合理设计结合位点 预定的特异性。 在该项目的后期阶段，VH 和 VL 基因将被改变为 重塑原始的 Fv 结合位点。 此外，单一多肽 Fv结构的结合位点将通过计算设计 并通过重组 DNA 方法执行。 这个的单一基因 结合位点是与细胞毒性剂基因融合的理想选择，例如 蓖麻毒素A亚基。 我们将生成这样一个嵌合物种进行试验 体外研究。

项目成果

A bifunctional fusion protein containing Fc-binding fragment B of staphylococcal protein A amino terminal to antidigoxin single-chain Fv.

• DOI: 10.1021/bi00487a005
• 发表时间: 1990-09
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: M. Tai;Meredith Mudgett-Hunter;Douglas Levinson;Gay‐May Wu;Edgar Haber;Hermann Oppermann;James S. Huston