Alfthan H, Stenman U H
Department of Obstetrics and Gynecology, Helsinki University Central Hospital, Finland.
J Clin Endocrinol Metab. 1990 Mar;70(3):783-7. doi: 10.1210/jcem-70-3-783.
We have developed a time-resolved immunofluorometric assay (IFMA) for quantification of the core fragment of the beta-subunit (c beta hCG) of hCG. The assay uses two monoclonal antibodies. One antibody is immobilized onto the wall of a microtiter strip well, and the other one is labeled with a europium chelate. The assay is sensitive (0.44 pmol/L = 4.6 ng/L), but cross-reaction with free beta-subunits of hCG (beta hCG) prevents direct determination of c beta hCG in serum. To circumvent this limitation we separated hCG and beta hCG from c beta hCG by gel chromatography and quantified each component in the fractions by specific IFMAs. The high sensitivity of the newly developed IFMA enabled us to demonstrate that serum from pregnant women contains c beta hCG and elutes at the same position in gel chromatography as c beta hCG purified from urine. The proportion of c beta hCG to hCG in pregnancy serum was 0.012-0.045% (mean +/- SD, 0.028 +/- 0.01) on a molar basis. Our finding of c beta hCG in serum confirms earlier reports suggesting that proteolytic degradation of beta hCG in the kidneys may not be the only pathway by which c beta hCG is formed.
我们开发了一种时间分辨免疫荧光分析法(IFMA),用于定量检测人绒毛膜促性腺激素(hCG)β亚基的核心片段(cβhCG)。该分析方法使用两种单克隆抗体。一种抗体固定在微量滴定板孔壁上,另一种用铕螯合物标记。该分析方法灵敏度高(0.44 pmol/L = 4.6 ng/L),但与hCG的游离β亚基(βhCG)发生交叉反应,妨碍了血清中cβhCG的直接测定。为克服这一局限性,我们通过凝胶色谱法将hCG和βhCG与cβhCG分离,并通过特异性IFMA对各组分进行定量。新开发的IFMA具有高灵敏度,使我们能够证明孕妇血清中含有cβhCG,并且在凝胶色谱中的洗脱位置与从尿液中纯化的cβhCG相同。在摩尔基础上,妊娠血清中cβhCG与hCG的比例为0.012 - 0.045%(平均值±标准差,0.028±0.01)。我们在血清中发现cβhCG,证实了早期报告,表明肾脏中βhCG的蛋白水解降解可能不是形成cβhCG的唯一途径。