Selective elution of target protein from affinity resins by a simple reductant with a thiol group.
We have made a chance discovery of selective elution of a specific binding protein from affinity resins by mixing them with aqueous solutions of a widely used reductant, 2-mercaptoethanol (2ME), under mild conditions. Our studies suggest this phenomenon would be generic, and could be a powerful method for identification of a specific binding protein. We here exhibit the experimental conditions and successful examples in which target proteins of benzensulfonamide and FK506 were selectively eluted from affinity resins bearing these compounds, while non-specific ones remained.
Department of Pharmacy, Hyogo University of Health Sciences, Chuo-ku, Kobe, Japan.
Miyuki
M
Mabuchi
Masayuki
M
Haramura
Tadashi
T
Shimizu
Tomoyuki
T
Nishizaki
Akito
A
Tanaka
D024402:Carbonic Anhydrase II / Q000737:chemistry / Q000302:isolation & purification; D002846:Chromatography, Affinity; D008623:Mercaptoethanol / Q000737:chemistry*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D019163:Reducing Agents / Q000737:chemistry*; D012117:Resins, Synthetic / Q000737:chemistry*; D013449:Sulfonamides / Q000737:chemistry; D016559:Tacrolimus / Q000737:chemistry; D022061:Tacrolimus Binding Protein 1A / Q000737:chemistry / Q000302:isolation & purification
D011506:Proteins; D019163:Reducing Agents; D012117:Resins, Synthetic; D013449:Sulfonamides; D008623:Mercaptoethanol; D024402:Carbonic Anhydrase II; D022061:Tacrolimus Binding Protein 1A; D016559:Tacrolimus