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识别在人类结肠癌与正常成人结肠组织中差异表达的癌胚抗原表位的单克隆抗体。

Monoclonal antibody recognizing a carcinoembryonic antigen epitope differentially expressed in human colonic carcinoma versus normal adult colon tissues.

作者信息

Zhang H Z, Ordonez N G, Batsakis J G, Chan J C

机构信息

Department of Tumor Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.

出版信息

Cancer Res. 1989 Oct 15;49(20):5766-73.

PMID:2477148
Abstract

Murine hybridomas were generated against purified carcinoembryonic antigen (CEA), and their monoclonal antibodies (MAb) were assayed for their ability to recognize specific CEA epitopes. One of the MAb, designated 7F, was found to recognize an epitope of CEA that was expressed in human colonic carcinoma tissues but not in normal colonic tissues. When extracts of 13 colonic carcinoma tissues and 9 normal colonic tissues were examined with the Western blot technique using MAb 7F, 11 colonic carcinoma tissues (85%) reacted with 7F, but none of the 9 normal colonic tissues (including many obtained from the areas adjacent to the colonic carcinomas) did. Western blot analysis indicated a Mr 180,000 band in 11 of 13 (85%) colonic carcinoma extracts. The limit of detectability of CEA was 0.5 micrograms/lane. According to immunohistochemical techniques, 16 of 18 (89%) formalin-fixed paraffin-embedded sections of colonic carcinomas reacted with MAb 7F, whereas 9 of 9 (100%) frozen sections of colonic carcinomas reacted with the antibody. Of the 32 paraffin-embedded and frozen sections of normal colonic tissues examined, none showed any reactivity with MAb 7F. MAb 7F did not react with nonspecific cross-reacting antigen either. This antibody has been examined for its usefulness in detecting CEA in suspension and has been found to work both in sandwich enzyme-linked immunosorbent assays and in sandwich radioimmune assays. The detection of CEA in colon tumors but not in normal colonic tissues may be attributed to two possible explanations. It is possible that the level of CEA expression in normal colonic tissue is below the sensitivity of the assays employed. Alternatively, MAb 7F may recognize a "specific" epitope of CEA which was found in colon tumors but not in CEA of normal colonic tissues. At the present time, it is not possible to discern between these two possibilities. Nevertheless, MAb 7F was capable of detecting the differential expression of CEA in colonic carcinomas and, therefore, may be useful for the immunodiagnosis, radioimaging, and immunotherapy of colon cancer.

摘要

制备了针对纯化癌胚抗原(CEA)的小鼠杂交瘤,并检测了它们的单克隆抗体(MAb)识别特定CEA表位的能力。其中一种单克隆抗体命名为7F,它能识别在人结肠癌组织中表达但在正常结肠组织中不表达的CEA表位。当用单克隆抗体7F通过蛋白质印迹技术检测13份结肠癌组织提取物和9份正常结肠组织提取物时,11份(85%)结肠癌组织与7F发生反应,但9份正常结肠组织(包括许多取自结肠癌旁区域的组织)均未发生反应。蛋白质印迹分析表明,13份结肠癌提取物中有11份(85%)出现了分子量为180,000的条带。CEA的可检测下限为0.5微克/泳道。根据免疫组织化学技术,18份福尔马林固定石蜡包埋的结肠癌切片中有16份(89%)与单克隆抗体7F发生反应,而9份结肠癌冰冻切片中有9份(100%)与该抗体发生反应。在检测的32份正常结肠组织石蜡包埋和冰冻切片中,均未显示与单克隆抗体7F有任何反应性。单克隆抗体7F也不与非特异性交叉反应抗原发生反应。已检测了该抗体在检测悬浮液中CEA方面的实用性,发现它在夹心酶联免疫吸附测定和夹心放射免疫测定中均有效。在结肠肿瘤中能检测到CEA而在正常结肠组织中检测不到,可能有两种解释。一种可能是正常结肠组织中CEA的表达水平低于所用检测方法的灵敏度。另一种可能是单克隆抗体7F可能识别的是在结肠肿瘤中发现但在正常结肠组织的CEA中不存在的CEA“特异性”表位。目前,无法区分这两种可能性。然而,单克隆抗体7F能够检测结肠癌中CEA的差异表达因此可能对结肠癌的免疫诊断、放射成像和免疫治疗有用。

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