Taguchi H, Miyamoto K, Takahashi Y, Ibuki Y, Igarashi M
Nihon Sanka Fujinka Gakkai Zasshi. 1985 Jun;37(6):973-81.
The ionic heterogeneity of prolactin (PRL) secreted in the human amniotic fluid was investigated using new precise chemical techniques. From five liters of amniotic fluid, the PRL fractions were purified with 70% ammonium sulfate precipitation, ethanol fractionation, desalting by gel filtration on Sephadex G-25, and ion-exchange chromatography using Mono Q on Fast Protein Liquid Chromatography (FPLC). The last FPLC was repeated until homogeneous preparations were obtained. Hormonal activity of each fraction was determined with a NIADDK radioimmunoassay (RIA) kit for human PRL. The PRL activity of the finally-purified component was also assayed by radioreceptor analysis (RRA) using rat liver cell membranes as the receptor. From the amniotic fluid, seven components of the PRL were eluted at NaCl concentrations from 0.065M to 0.13M in 0.02M Tris-HCl, pH 8.0. The RRA/RIA values of these seven components were different from each other (from 0.32 to 0.97). The lower the NaCl concentrations, the higher the RRA/RIA values were. The molecular weight of each component determined by gel filtration on TSK-SW 3000 with HPLC, was similar (MW 23,000) corresponding to "small-PRL".
利用新的精密化学技术研究了人羊水中分泌的催乳素(PRL)的离子异质性。从5升羊水中,通过70%硫酸铵沉淀、乙醇分级分离、在Sephadex G-25上进行凝胶过滤脱盐以及在快速蛋白质液相色谱(FPLC)上使用Mono Q进行离子交换色谱法来纯化PRL组分。重复最后一步FPLC操作,直到获得均一的制剂。使用NIADDK人PRL放射免疫分析(RIA)试剂盒测定每个组分的激素活性。最终纯化组分的PRL活性也通过以大鼠肝细胞膜为受体的放射受体分析(RRA)进行测定。在0.02M Tris-HCl(pH 8.0)中,从羊水中洗脱得到7种PRL组分,其NaCl浓度范围为0.065M至0.13M。这7种组分的RRA/RIA值彼此不同(从0.32至0.97)。NaCl浓度越低,RRA/RIA值越高。通过HPLC在TSK-SW 3000上进行凝胶过滤测定的每个组分的分子量相似(MW 23,000),对应于“小分子PRL”。