Concentration of 17beta-estradiol using an immunoaffinity porous hollow-fiber membrane.
We describe attempts to achieve high throughput of 17beta-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 microg/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 microg 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.
Nishiyama
S
S
Department of Materials Technology, Chiba University, Japan.
Goto
A
A
Saito
K
K
Sugita
K
K
Tamada
M
M
Sugo
T
T
Funami
T
T
Goda
Y
Y
Fujimoto
S
S
D000465:Algorithms; D004797:Enzyme-Linked Immunosorbent Assay; D004958:Estradiol / Q000032:analysis* / Q000276:immunology; D007120:Immunochemistry; D008024:Ligands; D008567:Membranes, Artificial; D010539:Permeability
D008024:Ligands; D008567:Membranes, Artificial; D004958:Estradiol