ENGINEERING OF ANTIBODY BINDING SITES TO TUMOR ANTIGENS

将制定基本方法和遗传要素， 生物合成抗体结合结构域的可互换特异性。 VH 基于抗地高辛抗体的Fv区的VL结构域将被 被改造成具有抗蛋白的高变片段 抗体的 这旨在改变其特异性，而无需 改造Fv的恒定框架。 这两种不同鼠的杂交种 单克隆结合区将在原核和真核细胞中表达 宿主以允许纯化复性的VH、VL和重组Fv。 对其绑定属性的分析将确定它是否完全起作用 如果没有，计算和物理化学研究将 为重新设计提供基础。 类似地，人Fv区的恒定框架将用于 支持鼠抗蛋白单克隆抗体的结合位点。 等 链内、种间杂交可以为人类提供独特优势， 疗法 特别地，最小化人对免疫缺陷病毒的免疫反应是一种有效的方法。 鼠结合位点是理想的，利用鼠单克隆抗体也是理想的 实验上可获得的结构特征。 尝试将 使VH和VL基因通过接头融合成单个Fv基因， 与组分的正常重折叠和重组相容 域. 该单一多肽链结合位点将可用于 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.