Immobilization of proteins on aldehyde-activated polyacrylamide supports

✍ Scribed by Morris B. Fiddler; Gary R. Gray

• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 560 KB
• Volume: 86
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(78)90799-6

✦ Synopsis

A method has been developed for the immobilization of proteins on derivatized polyacrylamide gels. Aminoethyl Bio-Gel P-I50 was converted to its stable N-2,3dihydroxypropyl derivative by borohydride reduction of the Schiff base formed with glyceraldehyde. Periodate oxidation of the modified gel provided a reactive aldehyde. which was subsequently coupled to protein by reductive amination with sodium cyanoborohydride. Coupling efficiencies were found to be >90% for concanavalin A and bovine serum albumin. and the gels contained as much as 5 and 20 mg of protein/ml of gel, respectively. Immobilized concanavalin A retained 89% of its binding capacity and was demonstrated to be chemically stable with variations in pH, and changes in concentrations of Triton X-100 and sodium dodecyl sulfate (at concentrations <O. 1%). Bovine P-hexosaminidase and P-glucuronidase, higher molecular weight proteins, were also bound with retention of activity, but with less efficiency. This procedure provides an efficient method for the covalent immobilization of proteins.