MOLECULAR IMMUNOLOGY

重组DNA技术极大地提高了我们的研究能力 基因结构和功能。这项技术现在可以应用于 生产单抗(Mab)。单抗有助于 蛋白质高度特异性识别的生化分析 一个单一的表位。生成单抗的传统方法不是 能够有效地检测对给定抗体的诱导抗体反应 抗原。例如，一只动物有5-10,000个不同的B细胞 克隆体，每个克隆都能产生针对一种抗原的独特抗体。 然而，利用目前的细胞融合技术，只有几百个 可以产生不同的抗体。重组DNA技术允许 待扩增的免疫球蛋白可变区基因，提供 用于使用噬菌体产生大的cDNA文库 Lambda-免疫-ZAP表达载体。这个库更容易 比细胞融合产生的杂交瘤更容易获得。筛查也是 大大增强，考虑到免疫球蛋白基因产品至少 一次可轻松检测50,000个克隆或1-10万个抗体 与筛选杂交瘤相比，杂交瘤是劳动密集型的， 既耗时又昂贵。一旦生产出来，这些表达载体 可被导入哺乳动物细胞或用于进行转基因 老鼠。 目前，我们已经克隆了PAN-ETS的重链和轻链基因 (T-7)针对人ETS2癌蛋白的抗体。那盏灯 链已成功插入lambda-LCL表达式 并构建了文库。工作继续进行，取得了进展 构建重链文库。

Recombinant DNA technology has greatly increased our ability to study gene structure and function. This technology can now be applied to monoclonal antibody (Mab) production. Monoclonal antibodies facilitate biochemical analysis of proteins by their highly-specific recognition of a single epitope. Conventional methods for generating Mabs are not capable of efficiently surveying the induced antibody response to a given antigen. For example, an individual animal has 5-10,000 different B-cell clones, each capable of producing unique antibodies to an antigen. However, with the current cell-fusion technique, only a few hundred different antibodies can be produced. Recombinant DNA technology allows the immunoglobin variable region genes to be amplified, which provides for the generation of a large CDNA library using the bacteriophage lambda-immuno-zap expression vectors. This library is much easier to access than hybridomas produced from cell fusion. Screening is also greatly enhanced, considering immunoglobin gene products of at least 50,000 clones or 1-10,000,000 antibodies can be readily examined in one day, compared to screening hybridomas, which is labor-intensive, time-consuming and expensive. Once produced, these expression vectors can be transfected into mammalian cells or used for making transgenic mice. Currently, we have cloned the heavy and light chain genes of the pan-ets (T-7) antibody directed against the human ETS2 oncoprotein. The light chain has been success-fully inserted into the lambda-Lcl expression vector and a library has been constructed. Work continues with progress to construct the heavy chain library.