单克隆抗体mab.ppt

* ? * * Figure 9-23. Organization of the k light-chain locus in human germ-line DNA. In a library spread over hundreds of kilobases of DNA, there are about 100 Vk segments, each with a unique sequence. The Vk segments occur in both transcriptional orientations, whereas the five Jk segments and one Ck segment are all in the 5 3 orientation (left to right in the illustration). One Lk-Vkunit is shown expanded. It consists of a promoter at the 5 end (beginning of transcription), an exon encoding a leader peptide (Lk), an intron, an exon encoding the Vk region, and a recognition signal. This signal guarantees that the Vk segment will join to a Jk segment. The arrow shows the orientation of the recognition site. See text for further discussion. [After Paul D. Gottleib, Mol. Immunol. 17:1423.] * Figure 9-25. Organization of heavy-chain gene segments in human germ-line DNA. The heavy-chain locus contains a library of D segments in addition to multiple V and J segments analogous to those in the k light-chain locus. The VH, D, and JH segments have recognition signals (red) that assure proper joining. The small arrows indicate the orientation of the recognition signals. By processes similar to those depicted in Figure 9-24, a D segment first joins to a JH segment with theintervening DNA being deleted; a VH segment then joins the preformed DJH unit, forming the variable region of the heavy-chain gene. Subsequent transcription yields a primary transcript, which is spliced to yield heavy-chain mRNA. The Cm segment encodes the constant region of the m heavy chain, which is found in IgM. Other C segments downstream from Cm (not shown) encode the constant regions of heavy chains in the other Ig classes. * * ? * * * * * * * * 1 * * * * 抗体库技术产生的三项技术基础 ? RT-PCR：能够克隆全套抗体可变区基因 ? 抗体基因片段在大肠杆菌的功能性表达 ? 噬菌体展示技术（phage display) 噬菌体表面展示技术(Phage display technology) 噬菌体表面展示技术即用PCR技术从生物体内扩增出整套编码人抗体的基因序列，克隆到噬菌体载体上，并以融合蛋白的形式表达到噬菌体表面，从而可以方便地利用抗原一抗体特异性结合进行筛选、扩增。 噬菌体抗体库技术 将从人或小鼠B淋巴细胞中分离的mRNA逆转录成cD