Xiang J H, Roder J, Hozumi N
Samuel Lunenfeld Research Institute of Mount Sinai Hospital, Toronto, Ontario, Canada.
Mol Immunol. 1990 Aug;27(8):809-17. doi: 10.1016/0161-5890(90)90091-d.
A mouse/human chimeric B72.3-1-3 antibody was produced by construction of a novel expression vector mpSV2neo-EP1-V-Cr1. This vector contains the neo gene as a selection marker, the murine immunoglobulin heavy chain promoter and enhancer, the murine V region cDNA containing mRNA splicing joint sequences, amplified and cloned by the PCR technique directly from the B72.3 hybridoma RNA, and the human genomic Cr1 region. The expression vector containing the murine/human chimeric immunoglobulin heavy chain gene was transfected into heavy chain loss mutant cell line, B72.3Ml. Chimeric B72.3-1-3 antibody was produced at 2 micrograms/ml and retained full binding reactivity to TAG72 compared to the murine B72.3 parental antibody. Using this method, chimeric immunoglobulin molecules can be produced rapidly in comparison with the cDNA and genomic cloning techniques.
通过构建新型表达载体mpSV2neo-EP1-V-Cr1产生了一种小鼠/人嵌合B72.3-1-3抗体。该载体包含作为选择标记的neo基因、小鼠免疫球蛋白重链启动子和增强子、含有mRNA剪接接头序列的小鼠V区cDNA（通过PCR技术直接从B72.3杂交瘤RNA扩增并克隆）以及人基因组Cr1区。将含有小鼠/人嵌合免疫球蛋白重链基因的表达载体转染到重链缺失突变细胞系B72.3M1中。嵌合B72.3-1-3抗体的产量为2微克/毫升，与小鼠B72.3亲本抗体相比，对TAG72保留了完全的结合反应性。与cDNA和基因组克隆技术相比，使用这种方法可以快速产生嵌合免疫球蛋白分子。
