STRUCTURAL REQUIREMENTS OF ANTIBODY COMBINING SITES

将用模型抗体进行研究，以排除这种贡献 抗体可变区的不同部分与抗原结合。 一系列实验将确定结构要求 通过氨基酸序列的逐步转换进行的高亲和力结合 一种低亲和力小鼠抗偶氮苯砷酸(Ars)抗体的研制 变成更高亲和力的序列。另一系列实验 将涉及到交换鼠标的可变区的部分 抗Ars抗体与鼠抗α(1-6)葡聚糖抗体的抗体。 这将包括检验假设，即高变区 单独携带抗体特异性，并将其插入到 另一个框架。上述实验将使用SITE完成 定向体外诱变克隆抗体基因，并引入 将这些基因导入适当的骨髓瘤或杂交瘤细胞系。变种人 产生的抗体将被测试抗原结合的特异性和 亲和力和使用标准免疫学的独特型表达 方法：研究方法。这些研究将增加我们对结构的理解 对抗体特异性和亲和力的要求，以及对结构的要求- 蛋白质的功能关系和折叠。他们会提供帮助的 培养设计特制抗体的知识，以供研究和 出于医疗目的。

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.