STRUCTURAL REQUIREMENTS OF ANTIBODY COMBINING SITES

将用模型抗体进行研究， 抗体可变区的不同部分与抗原结合。 一系列的实验将确定的结构要求， 通过氨基酸序列的逐步转化的高亲和力结合 针对偶氮苯胂酸盐（Ars）的低亲和力小鼠抗体 插入到更高亲和力的序列中。 另一系列实验 将涉及互换小鼠可变区的部分 抗Ars抗体与小鼠抗α（1-6）葡聚糖抗体的那些。 这将包括测试假设，高变区 单独携带抗体特异性，并且当插入到 另一个框架。 上述实验将使用现场 克隆抗体基因的体外定向诱变，以及引入 将这些基因导入适当的骨髓瘤或杂交瘤细胞系中。 突变 将测试产生的抗体的抗原结合特异性， 亲和力和独特型表达，使用标准免疫学方法 方法. 这些研究将增加我们对结构的理解。 抗体特异性和亲和力的要求，以及结构的要求- 蛋白质的功能关系和折叠。 他们会帮助 发展知识来设计用于研究的定制抗体， 医疗目的。

Studies will be undertaken with model antibodies to disect the contribution of different portions of the antibody variable regions to antigen binding. One series of experiments will determine the structural requirements for high affinity binding by a stepwise conversion of the amino acid sequence of a lower affinity mouse antibody directed against azophenylarsonate (Ars) into the sequence of a higher affinity one. Another series of experiments will involve interchanging portions of the variable regions of a mouse anti-Ars antibody with those of a mouse anti-Alpha(1-6) dextran antibody. This will include testing the hypothesis that the hypervariable regions alone carry the antibody specificity and will retain it when inserted into another framework. The above experiments will be accomplished using site directed in vitro mutagenesis of cloned antibody genes, and introduction of these genes into appropriate myeloma or hybridoma cell lines. The mutant antibodies produced will be tested for antigen binding specificity and affinity and for idiotypic expression using standard immunological methods. These studies will increase our understanding of the structural requirements for antibody specificity and affinity, and of the structure - function relationship and folding of proteins in general. They will help develop the knowledge to design tailor - made antibodies for research and medical purposes.