Raam S, Vrabel D M
Clin Chem. 1986 Aug;32(8):1496-502.
Using an enzyme immunoassay (EIA) procedure, we evaluated the ability of the glass beads in the ER-EIA kit of Abbott Laboratories, which were coated with monoclonal anti-estrogen receptor (ER) antibodies, to bind hormone-free and hormone-filled Type I ER in cytosols, buffer washes, and 0.4 mol/L KCI extracts of ultracentrifugal pellets of breast cancer tissue homogenates. The unmodified ER-EIA technique yields higher values than does the dextran-coated charcoal (DCC) method for Type I ER. However, the antibody-coated beads fail to bind hormone-free ER and react with only a certain proportion of ER-[3H]estradiol complexes. The antigen saturation limit of the beads could not be determined because the quantity of antigen bound by the beads was disproportionate and unrelated to the total amount of the antigen available in the cytosols. In buffer extracts of tissue pellets that were ER-negative by the DCC assay, the EIA method detected high quantities of ER. We recommend checking the ability of the monoclonal antibodies to recognize proteins other than Type I ER in the extra-nuclear and nuclear compartments of target cells before using them for immunohistochemical detection of ER.
我们采用酶免疫测定（EIA）方法，评估了雅培实验室雌激素受体酶免疫测定（ER-EIA）试剂盒中包被有抗雌激素受体（ER）单克隆抗体的玻璃珠，在乳腺癌组织匀浆超速离心沉淀的胞质溶胶、缓冲液洗脱液和0.4mol/L KCl提取物中，与无激素和有激素的Ⅰ型ER结合的能力。对于Ⅰ型ER，未改良的ER-EIA技术比葡聚糖包被活性炭（DCC）法得出的值更高。然而，包被抗体的珠子无法结合无激素的ER，且仅与一定比例的ER-[³H]雌二醇复合物发生反应。由于珠子结合的抗原量不成比例且与胞质溶胶中可用的抗原总量无关，因此无法确定珠子的抗原饱和极限。在DCC检测为ER阴性的组织沉淀缓冲液提取物中，EIA方法检测到大量的ER。我们建议在将单克隆抗体用于ER的免疫组织化学检测之前，先检查其识别靶细胞胞质和细胞核区室中除Ⅰ型ER之外其他蛋白质的能力。
