Construction of Libraries of antibodies and its application

我们研究了构建抗体文库的条件，其中抗体只能通过各种抗原筛选才能分离出来。众所周知，抗体通过位于H链和L链V区的六个CRDs（互补性决定区）形成的结构特异性地与抗原结合。我们构建抗体文库的基本设计是使6个cdr的氨基酸序列多样化。我们使用了噬菌体展示系统。首先，我们比较了Fab型、单链Fv型和Fv型在噬菌体表面表达的抗体形式，我们采用Fab型抗体。接下来，我们开发了使用类固醇抗原和抗体改变抗原特异性的策略。第三，我们进行了cdr接枝实验，发现在框架区域的几个位置上的氨基酸及其组合对cdr形成的结构有很大的影响。因此，我们获得了抗原-抗体相互作用和噬菌体展示系统特性的各种宝贵信息。用于构建抗体文库。然而，我们后来采取了完全不同的策略。

We examined the conditions for constructing libraries of antibodies from which antibodies can be isolating only by screening with various antigens. It has been well known that antibodies bind specifically to antigens through the structures formed by six CRDs (complementarity-determining regions) that are located in V regions of H and L chains. Our basic design for construction of libraries of antibodies is diversification of amino acid sequences of six CDRs. We used the phage -display system. First, we compared the form of antibody expressed on the phage surface among Fab form, single chain Fv form and Fv form and we adopted the Fab form of antibody. Next, we have developed the strategy for changing antigen specificity using steroid antigens and antibodies. Thirdly, we performed CDR-grafting experiments and found that amino acids at several positions in the framework regions and their combinations gave substantial effects on the structures formed by the CDRs. Thus, we obtained various precious informations upon antigen-antibody interactions and the characteristics of phage-display system. For construction of libraries of antibodies. however, we adopted completely different strategies later on.