Protein immobilization on carbon nanotubes through a molecular adapter

A new approach to the modification of carbon nanotubes with biomolecules for the development of nanoscale biosensors is presented. Alkaline phosphatase was immobilized on the surface of multi-wall carbon nanotubes utilizing a layer-by-layer methodology. Carbon nanotubes were incubated with streptavidin, resulting in the formation of a protein layer on the surface of the nanotubes. Biotinylated alkaline phosphatase was then allowed to bind to streptavidin, anchoring the sensing protein onto the surface. Electrochemical biosensors were constructed by using carbon nanotubes compacted into pellets. 1-Naphthyl phosphate, which is hydrolyzed by alkaline phosphatase to the electroactive 1-naphthol, was used as a substrate. Electrodes constructed in this manner were observed to generate an electrochemical signal that was a function of substrate concentration.