Qi Y, Xiang J
Saskatoon Cancer Center, Department of Microbiology, University of Saskatchewan, Canada.
Hum Antibodies Hybridomas. 1995;6(4):161-6.
SP2/0Ag14 murine myeloma cells transfected with the expression vector mpSV2neo-EP-FV-CH2-3-PA containing the single gene FV-CH2-3 secreted a single-gene-encoded chimeric antibody molecule FV/M4. This single-chain protein consisted of the heavy- and light-chain variable (VH and VL) domains covalently joined through a flexible linker peptide, while the carboxyl end of VL domain was connected to the amino terminus of hinge region of the ccM4 heavy-chain. Our data showed that the FV/M4 retained both its immunoreactivity for tumor-associated TAG72 antigen and its cytolytic activity to tumor cells as did the parental ccM4 antibody. Therefore, this single-gene-construct approach circumvents inefficiencies inherent in delivering two genes into a mammalian cell for assembly of a functional chimeric antibody and provides an alternative for construction of chimeric antibodies. It is particularly attractive for ex vivo transfection of cells from patients for certain gene-therapy modalities not only for cancer but also for a range of diseases in which immunotherapeutic approaches are used.
用含有单基因FV-CH2-3的表达载体mpSV2neo-EP-FV-CH2-3-PA转染的SP2/0Ag14鼠骨髓瘤细胞分泌了单基因编码的嵌合抗体分子FV/M4。这种单链蛋白由通过柔性接头肽共价连接的重链和轻链可变区（VH和VL）组成，而VL结构域的羧基末端连接到ccM4重链铰链区的氨基末端。我们的数据表明，FV/M4保留了其对肿瘤相关TAG72抗原的免疫反应性以及对肿瘤细胞的细胞溶解活性，与亲本ccM4抗体相同。因此，这种单基因构建方法避免了将两个基因导入哺乳动物细胞以组装功能性嵌合抗体时固有的低效率，并为嵌合抗体的构建提供了一种替代方法。对于某些基因治疗方式，尤其是用于癌症以及一系列采用免疫治疗方法的疾病的患者细胞的离体转染，它特别具有吸引力。
