Okumura H, Sudo T, Fujisawa M, Namba S, Matsushima S
Nihon Naibunpi Gakkai Zasshi. 1976 Feb 20;52(2):105-13. doi: 10.1507/endocrine1927.52.2_105.
A crude hCG with an activity of about 3,000 IU per mg was purifed to 10,000--15,000 IU per mg of dry weight using Amberlite CG-50 chromatography combined with DEAE-Sephadex A-50 and Sephadex G-75. The alpha and beta subunits of hCG were prepared by urea-treatment of the hormone and isolated by DEAE-Sephadex A-50 chromatography. Further purification of the subunits was achieved by gel filtration on a Sephadex G-76 column. For radioimmunoassay hCG was iodinated by the DMSO-chloramine T method. Iodination of hCG with non-radioactive iodine revealed that the addition of DMSO to the iodination mixture seemed to reduce the iodination damage to the antigenic activity of the hormone. Non-radioactive iodine substituted hCG accomplished by the DMSO-chloramine T method showed 1.5 times more immunoreactive in the hCG radioimmunoassay than hCG iodinated by the usual chloramine T method. The radioimmunoassay of the hCG-beta subunit developed in our laboratory was satisfactory with respect to specificity; hLH, hFSH, hTSH and hCG-alpha subunit tested were cross-reacted very poorly in our assay system. Desialylated-hCG and subunits, whose biologic potency was almost zero, exhibited also decreased immunoreactivity, about 30% of the native hormones with grossly unimpaired parallelism in their respective homologous radioimmunoassays. The concentrations of hCG and the subunits were determined on human sera from pregnant patients during the course of pregnancy. The hCG levels reached to the peak at the first trimester of the pregnancy, however, the hCG-beta subunits varied in their concentrations poorly throughout the pregnancy periods. The hCG-alpha levels, on the other hand, depicted two distinct peaks, at the early period and the term of pregnancy.
使用Amberlite CG - 50色谱法结合DEAE - Sephadex A - 50和Sephadex G - 75，将每毫克活性约为3000国际单位的粗制人绒毛膜促性腺激素（hCG）纯化至每毫克干重10000 - 15000国际单位。通过对该激素进行尿素处理制备hCG的α和β亚基，并通过DEAE - Sephadex A - 50色谱法分离。通过在Sephadex G - 76柱上进行凝胶过滤进一步纯化亚基。对于放射免疫分析，hCG采用二甲基亚砜 - 氯胺T法进行碘化。用非放射性碘对hCG进行碘化显示，在碘化混合物中加入二甲基亚砜似乎能减少对激素抗原活性的碘化损伤。通过二甲基亚砜 - 氯胺T法完成的非放射性碘取代hCG在hCG放射免疫分析中的免疫反应性比用常规氯胺T法碘化的hCG高1.5倍。我们实验室开发的hCG - β亚基放射免疫分析在特异性方面令人满意；所测试的人促黄体生成素（hLH）、人促卵泡生成素（hFSH）、人促甲状腺激素（hTSH）和hCG - α亚基在我们的分析系统中交叉反应非常弱。去唾液酸化的hCG及其亚基的生物活性几乎为零，其免疫反应性也降低，在各自同源放射免疫分析中约为天然激素的30%，且平行性基本未受影响。在妊娠期间测定了孕妇血清中hCG及其亚基的浓度。hCG水平在妊娠早期达到峰值，然而，hCG - β亚基在整个孕期的浓度变化不大。另一方面，hCG - α水平在妊娠早期和足月时出现两个明显峰值。
