Reversible and irreversible immobilization of enzymes on Graphite Fibrils

Graphite Fibrils are hollow tubes (0.01 x 1-10 microns) consisting of concentric layers of graphite. Fibrils can be chemically modified to introduce surface functionalities such as carboxyl groups. Carboxylated fibrils were further functionalized to amino alkyl derivatives which were covalently linked to an enzyme, horseradish peroxidase (HRP). HRP fibrils showed substantial catalytic activity. Additionally, carboxyl fibrils were derivatized with specific inhibitors of the enzymes beta-galactosidase (beta-Gal) and alkaline phosphatase (AP). Enzyme inhibitor-modified fibrils were used to specifically purify both beta-Gal (using beta-Gal inhibitor fibrils) and AP (using AP inhibitor fibrils) from mixtures of these two enzymes. These studies demonstrate the feasibility of using Graphite Fibrils as supports in biocatalysis and biospecific affinity chromatography.