ENGINEERING OF ANTIBODY BINDING SITES TO TUMOR ANTIGENS

将开发用于生产的基本方法和遗传要素 具有可互换特异性的生物合成抗体结合域。VH 而基于抗地高辛抗体Fv区的VL结构域将是 重组以获得抗蛋白质的高可变片段 抗体。这是为了改变其特殊性，而不需要 重塑FV的恒定框架。这是两只不同小鼠的杂交 克隆结合区将在原核和真核细胞中表达 允许纯化复性的VH、VL和重组FV的宿主。 对其绑定属性的分析将确定它是否完全起作用 按计划进行，如果没有，计算和物理化学研究将 为重新设计提供了依据。 类似地，人类FV区域的恒定框架将用于 支持小鼠抗蛋白单抗的结合部位。是这样的 链内、种间杂交可能为人类提供独特的优势 心理治疗。特别是，最大限度地减少人类对 小鼠结合部位是可取的，利用小鼠单抗也是可取的 可通过实验访问的结构特征。尝试次数将是 通过连接子将VH和VL基因融合成单个FV基因 兼容正常的元件重折叠和重组 域名。这个单一的多肽链结合位点将用于 生物活性蛋白的免疫靶向。

Basic methodology and genetic elements will be developed for production of biosynthetic antibody binding domains of interchangeable specificity. VH and VL domains based upon the Fv region of an anti-digoxin antibody will be refashioned to have the hypervariable segments of an anti-protein antibody. This is intended to alter its specificity without the need to remake constant framework of the Fv. This hybrid of two different murine monoclonal binding regions will be expressed in prokaryotic and eukaryotic hosts to permit purification of renatured VH,VL, and recombined Fv. Analysis of its binding properties will determine if it functions entirely as planned, and if not, computational and physicochemical studies will provide the basis for redesign. Similarly, the constant framework of a human Fv region will be used to support the binding site from a murine anti-protein monoclonal. Such intrachain, interspecies hybrids could offer unique advantages for human therapy. In particular, minimization of a human immune reaction to a murine binding site is desirable, as is utilization of murine monoclonal structural features which are experimentally accessible. Attempts will be made to fuse VH and VL genes into a single Fv gene via a linker which will be compatible with normal refolding and recombination of component domains. This single polypeptide chain binding site would be useful for immunotargeting of biologically active proteins.