1. Concentration of 17[beta]-estradiol using an immunoaffinity porous hollow-fiber membrane
- Author
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Nishiyama, S., Goto, A., Saito, K., Sugita, K., Tamada, M., Sugo, T., Funami, T., Goda, Y., and Fujimoto, S.
- Subjects
Estradiol ,Membranes (Technology) -- Usage ,Immunoenzyme technique -- Methods ,Chemistry - Abstract
We describe attempts to achieve high throughput of 17[beta]-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 [micro]g/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 [micro]g was quantitatively eluted with 3-5 mL of methanol in the permeation mode. The higher permeation rate of the E2 solution resulted in the higher overall binding rate of E2 to the anti-ES-antibody-immobilized porous hollow-fiber membrane because of the negligible diffusional mass-transfer resistance of E2 to the antibody.
- Published
- 2002