Hardman N, Gill L L, De Winter R F, Wagner K, Hollis M, Businger F, Ammaturo D, Buchegger F, Mach J P, Heusser C
Sections of Biotechnology, CIBA-GEIGY, Basal, Switzerland.
Int J Cancer. 1989 Sep 15;44(3):424-33. doi: 10.1002/ijc.2910440308.
A procedure was devised for the identification and specific cloning of functionally rearranged variable region immunoglobulin (Ig) gene segments from genomic DNA of a murine hybridoma cell line which produces a high-affinity monoclonal antibody (MAb) directed against human carcinoembryonic antigen (CEA). The cloned, functionally-rearranged murine Ig H-chain and L-chain variable region gene segments were incorporated into plasmid vectors capable of directing the expression of a chimaeric mouse-human antibody molecule with human (gamma 4, kappa) constant region sequences. Expression plasmids were transfected into a mouse myeloma cell line by electroporation and transfectomas secreting functional chimaeric antibody selected. Chimaeric antibody generated by transfectomas was analysed and shown to compete effectively with its murine counterpart for binding to the CEA epitope, and to have an equivalent antigen-binding affinity. This anti-CEA recombinant antibody should find application in in vivo diagnosis by immunoscintigraphy of human colonic carcinoma, and possibly also in therapy of the disease, overcoming some of the difficulties associated with the repeated use of non-human immunoglobulins in human patients.
设计了一种程序，用于从产生针对人癌胚抗原（CEA）的高亲和力单克隆抗体（MAb）的鼠杂交瘤细胞系的基因组DNA中鉴定和特异性克隆功能重排的可变区免疫球蛋白（Ig）基因片段。将克隆的、功能重排的鼠Ig重链和轻链可变区基因片段整合到能够指导表达具有人（γ4，κ）恒定区序列的嵌合鼠-人抗体分子的质粒载体中。通过电穿孔将表达质粒转染到小鼠骨髓瘤细胞系中，并选择分泌功能性嵌合抗体的转染瘤。对转染瘤产生的嵌合抗体进行分析，结果表明其能与其鼠源对应物有效竞争结合CEA表位，且具有同等的抗原结合亲和力。这种抗CEA重组抗体应可用于人结肠癌的免疫闪烁体内诊断，也可能用于该疾病的治疗，克服了在人类患者中重复使用非人免疫球蛋白所带来的一些困难。
