cDNA sequence analysis of monoclonal antibody FU-MK-1 specific for a transmembrane carcinoma-associated antigen, and construction of a mouse/human chimeric antibody.
cDNA sequence analysis of monoclonal antibody FU-MK-1 specific for a transmembrane carcinoma-associated antigen, and construction of a mouse/human chimeric antibody.
复制标题
对跨膜癌相关抗原特异的单克隆抗体 FU-MK-1 进行 cDNA 序列分析，并构建小鼠/人嵌合抗体。
作者：
Fumiko Arakawa;Takafumi Yamamoto;H. Kanda;Takeshi Watanabe;Masahide Kuroki
针对人胃腺癌提出的小鼠单克隆抗体FU-MK-1，识别存在于大多数腺癌上的跨膜抗原GA733-2，似乎对这些癌症的免疫诊断和免疫治疗具有潜在的效用。然而，它们在体内应用的一个固有问题是人抗小鼠抗体反应。本研究采用逆转录-聚合酶链反应方法克隆了FU-MK-1重链和轻链（V(H)和Vkappa）可变区基因，并对其进行了测序。然后，我们将FU-MK-1的V(H)和Vkappa基因分别与人Cgamma1和Ckappa基因融合，并在哺乳动物细胞表达载体上将嵌合的H链和L链基因相互连接，构建鼠/人嵌合抗体，命名为Ch FU-MK-1。最终构建的基因通过电穿孔法转染到小鼠不产生igg的杂交瘤细胞中。由此制备的chfu - mk -1抗体与人腺癌细胞结合，并竞争性地抑制亲本FU-MK-1与腺癌细胞的结合。Ch FU-MK-1还显示出一种有效的抗体依赖细胞介导的细胞毒性（ADCC），以人外周血单个核细胞作为效应物来对抗腺癌细胞，这表明这种嵌合抗体似乎适合于体内治疗方法。
Mouse monoclonal antibody (MAb) FU-MK-1, raised against a human gastric adenocarcinoma, recognizes a transmembrane antigen, GA733-2, present on most adenocarcinomas and seems to be of potential utility for immunodiagnosis and immunotherapy of those cancers. However, an inherent problem in their in vivo application is the human anti-mouse antibody response. In this study, we cloned and sequenced the variable region genes of the heavy and light chains (V(H) and Vkappa) of FU-MK-1 using the reverse transcription-polymerase chain reaction method. Then, we constructed a mouse/human chimeric antibody, designated as Ch FU-MK-1, by fusing the FU-MK-1 V(H) and Vkappa genes to the human Cgamma1 and Ckappa genes, respectively, and by ligating the chimeric H and L chain genes to each other in a mammalian cell expression vector. The final gene construct was transfected into mouse non-Ig-producing hybridoma cells by electroporation. The Ch FU-MK-1 antibody thus prepared bound to human adenocarcinoma cells and competitively inhibited the binding of the parental FU-MK-1 to the adenocarcinoma cells. Ch FU-MK-1 also showed a potent antibody-dependent cell-mediated cytotoxicity (ADCC) with human peripheral blood mononuclear cells as effectors against the adenocarcinoma cells, indicating that this chimeric antibody seems to be suitable for in vivo therapeutic approaches.
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DOI：
10.1073/pnas.85.13.4852
发表时间：
1988-07-01
影响因子：
11.1
作者：
STEPLEWSKI, Z;SUN, LK;KOPROWSKI, H
通讯作者：
KOPROWSKI, H
DOI：
10.1073/pnas.87.9.3542
发表时间：
1990-05-01
影响因子：
11.1
作者：
SZALA, S;FROEHLICH, M;LINNENBACH, AJ
通讯作者：
LINNENBACH, AJ
DOI：
10.1073/pnas.81.21.6851
发表时间：
1984-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
MORRISON, SL;JOHNSON, MJ;OI, VT
通讯作者：
OI, VT
长期鼠单克隆抗体 17-1A 治疗胰腺癌的 II 期多中心评估。
DOI：
10.1097/00002371-199302000-00005
发表时间：
1993
期刊：
Journal of immunotherapy with emphasis on tumor immunology : official journal of the Society for Biological Therapy
影响因子：
--
作者：
Weiner,LM;Harvey,E;Padavic-Shaller,K;Willson,JK;Walsh,C;LaCreta,F;Khazaeli,MB;Kirkwood,JM;Haller,DG
通讯作者：
Haller,DG
由针对人类肿瘤相关抗原的鼠单克隆抗体表达的相同 D 区序列。
DOI：
--
发表时间：
1990
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Caton,AJ;Herlyn,D;Ross,AH;Koprowski,H
通讯作者：
Koprowski,H