Anti-IgG immobilized controlled-pore glass : thionyl chloride-activated succinamidopropyl-glass as a covalent immobilization matrix

• Published in: Applied biochemistry and biotechnology 1992-08, Vol.36 (2), p.87-96
• Main Authors: STABEL, T. J, CASALE, E. S, SWAISGOOD, H. E, HORTON, H. R
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Anti-Idiotypic; Biological and medical sciences; Biotechnology; Blotting, Western; Cattle; Chromatography, Affinity - methods; Enzyme-Linked Immunosorbent Assay; Fundamental and applied biological sciences. Psychology; gamma-Glutamyltransferase - isolation & purification; Glass; Immobilization of enzymes and other molecules; Immobilization techniques; Immunoglobulin G; Methods. Procedures. Technologies; Microspheres; Oxidoreductases - isolation & purification; Rabbits; Sulfur Oxides
• ISSN: 0273-2289; 1559-0291

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.