Production of monoclonal antibodies to human estrogen‐receptor protein (ER) using recombinant ER (RER)
Production of monoclonal antibodies to human estrogen‐receptor protein (ER) using recombinant ER (RER)
复制标题
使用重组 ER (RER) 生产抗人雌激素受体蛋白 (ER) 的单克隆抗体
DOI：
--
复制
发表时间：
1993
影响因子：
6.4
通讯作者：
J. Wafflart
中科院分区：
文献类型：
--
作者：
T. Saati;Edith Cohen‐Knafo;P. Caverivière;G. Delsol;S. Clamens;J. Faye;H. Prats;F. Bayard;J. Coindre;J. Wafflart
使用重组雌激素受体蛋白（RER）免疫BALB/c小鼠的脾细胞制备了两种单克隆抗体（MAb），IC 5和ID 5。在免疫印迹中，两种MAb均与通过转化E. coli中表达ER的质粒载体转染COS细胞。两种MAb的表位均位于受体的N末端结构域（A/B区）。在正常人体组织中，IC 5和ID 5与已知含有大量ER的细胞反应，例如乳腺和子宫的细胞。染色主要位于细胞核中，细胞质反应性很小或没有。IC 5和ID 5与通常认为ER阴性的组织不反应。在冷冻切片上使用免疫组织化学（IHC）方法进一步测试这2种MAbs对不同肿瘤类型的反应。在乳腺癌中，在冷冻切片上获得的结果与使用常规放射性配体葡聚糖包被活性炭（DCC）测定的结果之间发现了良好的相关性。还使用抗原修复法在常规处理的石蜡切片上评估了IC 5和ID 5 MAb的免疫染色。染色结果与几乎所有乳腺癌的冰冻切片结果相当。两种抗体对源自其他非雌激素依赖性器官的人肿瘤均获得阴性反应。因此，IC 5和ID 5单克隆抗体可能在常规诊断组织病理学中具有价值，用于评估人类癌症中的雌激素受体含量。
Two monoclonal antibodies (MAbs), IC5 and ID5, were produced using spleen cells from BALB/c mice immunized with recombinant estrogen‐receptor protein (RER). On immunoblotting, both MAbs reacted with the 67‐kDa polypeptide chain obtained by transformation of E. coli and transaction of COS cells with plasmid vectors expressing ER. The epitopes of both MAbs were in the N‐terminal domain (A/B region) of the receptor. In normal human tissues, IC5 and ID5 reacted with cells known to contain large amount of ER, such as cells of the mammary gland and the uterus. Staining was localized predominantly in nuclei with little or no cytoplasmic reactivity. IC5 and ID5 were unreactive with tissues usually considered to be negative for ER. The reactions of these 2 MAbs were further tested on different tumor types, using immunohistochemical (IHC) method on frozen sections. In breast cancer, a good correlation was found between the results obtained on frozen sections and those using the conventional radioligand dextran‐coated charcoal (DCC) assay. Immunostaining with IC5 and ID5 MAbs was also assessed on routinely processed paraffin sections using the antigen‐retrieval method. Staining was comparable to that obtained on frozen sections in virtually all the breast carcinomas. Negative reactions were consistently obtained with both antibodies on human neoplasms derived from other non‐estrogen‐dependent organs. IC5 and ID5 MAbs may thus be of value in routine diagnostic histopathology for assessment of the estrogen‐receptor content in human carcinomas.
DOI：
10.1016/0022-4731(84)90188-2
发表时间：
1984-01-01
影响因子：
4.1
作者：
GREENE, GL;SOBEL, NB;JENSEN, EV
通讯作者：
JENSEN, EV
DOI：
10.1073/pnas.77.9.5115
发表时间：
1980-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
GREENE, GL;NOLAN, C;JENSEN, EV
通讯作者：
JENSEN, EV
DOI：
10.1073/pnas.87.1.142
发表时间：
1990-01-01
影响因子：
11.1
作者：
SANO, T;CANTOR, CR
通讯作者：
CANTOR, CR