Covalent protein immobilization on glass surfaces: application to alkaline phosphatase.
Lyophilized alkaline phosphatase (ALPase) was immobilized on aminated glass surfaces using the in vacuo cross-linking process [Simons, B.L., King, M.C., Cyr, T., Hefford, M.A., Kaplan, H., 2002. Zero-length cross-linking of lyophilized proteins. Protein Sci. 11, 1558-1564]. In this procedure, amide bonds were formed between carboxyl groups on the protein and amino groups on the glass surface. After the non-covalently attached enzyme was removed the immobilized ALPase not only retained its activity but could also be used, washed and reused at least six times without significant loss of activity. An average of 1.4+/-0.6 mg of reusable ALPase per gram of glass fibre was immobilized based on the activity of the soluble equivalent.
Taylor
Russell H
RH
Department of Biochemistry, University of Ottawa, Canada K1H 8M5.
Fournier
Sylvie M
SM
Simons
Brigitte L
BL
Kaplan
Harvey
H
Hefford
Mary Alice
MA
D000469:Alkaline Phosphatase / Q000032:analysis / Q000737:chemistry*; D020099:Coated Materials, Biocompatible / Q000032:analysis / Q000737:chemistry*; D003460:Crystallization / Q000379:methods*; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000032:analysis / Q000737:chemistry; D005898:Glass / Q000737:chemistry*; D008422:Materials Testing; D011485:Protein Binding
D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D000469:Alkaline Phosphatase