Singleton T P, Niehans G A, Gu F, Litz C E, Hagen K, Qiu Q, Kiang D T, Strickler J G
Department of Laboratory Medicine and Pathology, University of Minnesota Hospital and Clinic, Minneapolis.
Hum Pathol. 1992 Oct;23(10):1141-50. doi: 10.1016/0046-8177(92)90032-x.
Commercially available monoclonal antibodies were tested for their ability to detect increased levels of c-erbB-2 protein in formalin-fixed, paraffin-embedded breast carcinomas. Of five antibodies studied, four (TAB-250, CB11, 3B5, and N3/D10) showed strong cytoplasmic membrane reactivity in 23% (11 of 47) of routinely processed tumors, although interpretation of the immunoreactivity with 3B5 and N3/D10 occasionally was difficult due to cytoplasmic granular staining. Since the c-erbB-2 oncogene is activated by DNA amplification and overexpression of mRNA and protein, the same tumors were analyzed for c-erbB-2 activation by other techniques. c-erbB-2 activation in these 11 tumors was confirmed by immunohistochemistry of frozen tissue (nine of nine tumors), in situ hybridization (nine of 11 tumors), and Southern blot analysis (five of eight tumors). In some of these tumors the failure to demonstrate c-erbB-2 DNA amplification may be due to the small percentage of malignant cells. One additional tumor showed probable c-erbB-2 protein overproduction based on strong immunoreactivity with two antibodies (TAB-250 and CB11), although no definite activation could be demonstrated by additional techniques. Three other tumors (6%) showed equivocal c-erbB-2 protein overproduction based on weak immunoreactivity only with TAB-250, although unequivocal activation could not be demonstrated by additional techniques. The 32 carcinomas (68%) that showed no significant immunoreactivity with any antibodies in routinely processed tissue also showed no detectable c-erbB-2 activation by additional techniques. We conclude that TAB-250 and CB11 are reliable antibodies for detecting c-erbB-2 protein overproduction in routinely processed tissue. TAB-250 also weakly stains a few tumors showing no definite c-erbB-2 activation by other techniques. Two additional antibodies (3B5 and N3/D10) detect c-erbB-2 protein overproduction in paraffin-embedded tissue, but are more difficult to interpret. A fifth antibody, TA-1, is an excellent reagent for use on frozen tissue, but prolonged formalin fixation may impair recognition of its antigenic epitope.
对市售单克隆抗体检测福尔马林固定、石蜡包埋乳腺癌中c-erbB-2蛋白水平升高的能力进行了测试。在所研究的5种抗体中，4种（TAB-250、CB11、3B5和N3/D10）在23%（47例中的11例）常规处理的肿瘤中显示出强烈的细胞质膜反应性，不过由于细胞质颗粒状染色，3B5和N3/D10的免疫反应性解释偶尔会有困难。由于c-erbB-2癌基因通过DNA扩增以及mRNA和蛋白质的过度表达而被激活，因此采用其他技术对相同肿瘤的c-erbB-2激活情况进行了分析。通过冷冻组织免疫组化（9例中的9例肿瘤）、原位杂交（11例中的9例肿瘤）和Southern印迹分析（8例中的5例肿瘤）证实了这11例肿瘤中的c-erbB-2激活。在其中一些肿瘤中，未能显示c-erbB-2 DNA扩增可能是由于恶性细胞所占比例较小。另一例肿瘤基于与两种抗体（TAB-250和CB11）的强免疫反应性显示可能存在c-erbB-2蛋白过量产生，不过通过其他技术未能证实明确的激活。另外3例肿瘤（6%）仅基于与TAB-250的弱免疫反应性显示c-erbB-2蛋白过量产生情况不明确，不过通过其他技术未能证实明确的激活。在常规处理组织中与任何抗体均无显著免疫反应性的32例癌（68%），通过其他技术也未检测到c-erbB-2激活。我们得出结论，TAB-250和CB11是检测常规处理组织中c-erbB-2蛋白过量产生的可靠抗体。TAB-250也会对一些通过其他技术未显示明确c-erbB-2激活的肿瘤进行弱染色。另外两种抗体（3B5和N3/D10）可检测石蜡包埋组织中的c-erbB-2蛋白过量产生，但更难解释。第五种抗体TA-1是用于冷冻组织的优良试剂，但长时间福尔马林固定可能会损害其对抗原表位的识别。
