Anti-IgG immobilized controlled-pore glass. Thionyl chloride-activated succinamidopropyl-glass as a covalent immobilization matrix.
Rabbit anti-bovine IgG was covalently immobilized on thionyl chloride-activated succinamidopropyl controlled-pore glass (CPG) beads (3000 A pore diam; 120/200 mesh). Thionyl chloride-activated beads remained stable for over 1 y retaining full capability of immobilizing protein upon recirculation of protein solution. The immobilized anti-bovine IgG is capable of binding IgG with a dissociation constant (Kd) of 9.45 x 10(-7) M and a capacity of 0.85 g/L of matrix. A column of immobilized anti-IgG is able to remove all detectable contaminating IgG from partially purified enzyme preparations of sulfhydryl oxidase and gamma-glutamyltransferase as determined by ELISA and Western blot. The column matrix could be regenerated by washing with 0.1M acetic acid, pH 2.8.
Stabel
T J
TJ
Department of Biochemistry, North Carolina State University, Raleigh 27695.
Casale
E S
ES
Swaisgood
H E
HE
Horton
H R
HR
D000818:Animals; D000888:Antibodies, Anti-Idiotypic; D015153:Blotting, Western; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005898:Glass; D007074:Immunoglobulin G; D008863:Microspheres; D010088:Oxidoreductases / Q000302:isolation & purification*; D011817:Rabbits; D013461:Sulfur Oxides; D005723:gamma-Glutamyltransferase / Q000302:isolation & purification*; D050862:Oxidoreductases Acting on Sulfur Group Donors
D000888:Antibodies, Anti-Idiotypic; D007074:Immunoglobulin G; D010088:Oxidoreductases; D013461:Sulfur Oxides; D005723:gamma-Glutamyltransferase; D050862:Oxidoreductases Acting on Sulfur Group Donors; C048844:anti-IgG; C023589:thionyl chloride