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.
Department of Biochemistry, North Carolina State University, Raleigh 27695.
T J
TJ
Stabel
E S
ES
Casale
H E
HE
Swaisgood
H R
HR
Horton
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