1992 Fiscal Year Final Research Report Summary
Preparation of Affinity Membrane for Protein Recovery
| Project/Area Number |
03650758
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
化学工学
|
|---|
| Research Institution | University of Tokyo |
|---|
Principal Investigator |
SAITO Kyoichi University of Tokyo Dept. of Chem. Eng., Associate Professor, 工学部, 助教授 (90158915)
|
|---|
| Project Period (FY) |
1991 – 1992
|
| Keywords | affinity / membrane / graft polymerization / radiation-induced / protein / hydrophobic amino acid / adsorption / elution curve |
|---|
| Research Abstract |
A porous hollow-fibre membrane containing L-phenylalanine as a pseudo-biospecific ligand has been prepared by radiation induced grafting of glycidyl methacrylate onto polyethylene microfiltration hollow fibre, followed by coupling of the produced epoxide group with L-phenylalanine. The remaining epoxide group was hydrolysed into a diol group with sulphuric acid. The L-phenylalanine and the diol group acted in a complementary way as a pseudo-biospecific ligand and a hydrophilic group, respectively. The adsorption capacity of bovine gamma globulin on the resulting porous adsorbent could be determined by the specific surface area of the adsorbent when the ligand was in excess over the protein.
A hollow-fibre affinity membrane containing hydrophobic amino acids as ligands was prepared by the radiation-induced grafting of glycidyl methacrylate onto a porous polyethylene hollow fibre and subsequent coupling with phenylalanine (Phe) or tryptophan (Trp). The densities of the Phe and Trp ligand of the resulting affinity membrane were 0.4 and 0.4 mol/kg, respectively. The Trp-containing affinity membrane exhibited a higher amount of adsorbed bovine y-globulin (BGG) than the Phe-containing membrane. To evaluate the adsorption behavior of the membrane, the BGG-containing buffer solution was permeated from the inside to the outside of the Trp-containing hollow-fibre affinity membrane through the ligand-immobilized pores. The breakthrough curves as a function of effluent volume coincided irrespective of the flow-rate, i.e. the residence time (55-220 s) of the solution across the membrane (thickness 0.83 mm), as a result of negligible mass transfer resistance. A series of chromatographic procedures, (adsorption-washing-elution) was repeated twice and a satisfactory quantitative elution was attained. The reproducible profile of the flux and the protein concentration assured a quantitative cycle of chromatography using the affinity membrane containing Trp as a ligand.
|
