MOLECULAR IMMUNOLOGY

LMO优化了寡核苷酸引物设计， 小鼠scFv克隆。 利用新设计的引物， 为了成功地克隆轻链和 抗ETS 1单克隆抗体E44重链及抗ETS 1小鼠 脾抗体 实验室目前正在努力解决 在轻链和重链组件中的问题， 通过更可预测的酶促连接进行有问题的PCR组装步骤。 在获得功能性抗ETS 1 scFv后， ETS 1细胞内功能的干扰。 表达 克隆的抗ETS 1单链抗体在真核细胞中的表达，以及结肠癌 细胞系，在ETS 1过表达后恢复正常， 锡永将尝试。 这些设计的scFv也将应用于 LMO内正在进行的各种ETS 1项目。 为了进一步探索小鼠scFv引物的用途，使用小鼠天然scFv引物， 为了便于表达和测试选择，开发了一个库 针对各种感兴趣的纯抗原和癌蛋白。 根据近年来的研究成果，建立了一种通用免疫PCR方法， 描述的程序，“免疫-PCR.“这是迄今为止最敏感的 方法可用于抗原检测，并可应用于各种 实验室和/或临床研究或诊断。

The LMO has optimized the oligonucleotide primer design for efficient mouse scFv cloning. With the newly-designed primers, it has been possible to successfully clone the variable domains of both the light chain and heavy chain of anti-ETS1 monoclonal antibody E44 and anti-ETS1 mouse spleen antibodies. The Laboratory is now making an effort to solve the problem in light chain and heavy chain assembly by replacing the problematic PCR assembly step by more predictable enzymatic ligation. After functional anti-ETS1 scFv's have been made, progress toward the goal of ETS1 intracellular function perturbation can be realized. Expression of the cloned anti-ETS1 scFv in eukaryotic cells, and a colon carcinoma cell line, which has been reverted back to normal after ETS1 overexpres- sion, will be attempted. Those designed scFv's will also be applied to various ETS1 projects ongoing within the LMO. To further explore the usage of the mouse scFv primers, a mouse naive scFv library has been developed in order to be expressed and test-selected against various pure antigens and oncoproteins of interest. A "Universal Immuno-PCR" method has been developed, based on the recently described procedure, "Immuno-PCR." This is, thus far, the most sensitive method available for antigen detection and can be applied to various laboratory and/or clinical studies or diagnosis.