Concentration of 17β-estradiol using an immunoaffinity porous hollow-fiber membrane
Concentration of 17β-estradiol using an immunoaffinity porous hollow-fiber membrane
复制标题
使用免疫亲和多孔中空纤维膜浓缩 17β-雌二醇
DOI：
10.1021/ac020141e
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发表时间：
2002
影响因子：
7.4
通讯作者：
S. Fujimoto
中科院分区：
文献类型：
--
作者：
S. Nishiyama;A. Goto;Kyoichi Saito;K. Sugita;M. Tamada;T. Sugo;T. Funami;Y. Goda;S. Fujimoto
我们描述了实现17β-雌二醇（E2）分析的高通量的尝试，包括使用多克隆抗体的免疫清洁膜和使用单克隆抗体的酶联免疫吸附测定（ELISA）的发展。将含环氧基团的单体甲基丙烯酸缩水甘油酯（GMA）接枝到多孔中空纤维膜上。随后，抗雌激素（ES）抗体，作为配体，与环氧基团偶联。配体密度范围为3.1至5.8毫克/克的GMA接枝的多孔中空纤维膜。迫使1.0 μg/L E2溶液以恒定渗透速率渗透通过由抗ES抗体固定的聚合物链镶边的孔。测定了穿透曲线，即，穿透中空纤维外部的流出物的E2浓度随流出物体积的变化而变化。在渗透模式下，用3 - 5 mL甲醇定量洗脱0.42 - 0.80 μg的结合E2。更高的渗透率。
We describe attempts to achieve high throughput of 17β-estradiol (E2) analysis, including the development of an immunocleanup membrane using polyclonal antibodies and an enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies. An epoxy-group-containing monomer, glycidyl methacrylate (GMA), was graft-polymerized onto a porous hollow-fiber membrane. Subsequently, anti-estrogen (ES) antibody, as a ligand, was coupled with the epoxy group. The ligand density ranged from 3.1 to 5.8 mg/g of the GMA-grafted porous hollow-fiber membrane. A 1.0 μg/L E2 solution was forced to permeate through pores rimmed by the anti-ES-antibody-immobilized polymer chains, at a constant permeation rate. A breakthrough curve, that is, the change in the E2 concentration of the effluent penetrating the outside of the hollow fiber with a change of the effluent volume, was determined. Bound E2 in amounts ranging from 0.42 to 0.80 μg was quantitatively eluted with 3−5 mL of methanol in the permeation mode. The higher permea...