3.2. Functionalization of electrode surface with biomolecules
For the detection of analytes from complex matrices, the transducer
element of the biosensor has to be coupled with a biorecognition
molecule, i.e. enzymes, antibodies, aptamers, etc., for
enhancing the biosensor selectivity and achieving high sensitivities
without using labeling or amplification techniques. The two general
approaches for modification of the electrode surface with
biomolecules are: covalent and non-covalent functionalization.
For covalent attachment of biomolecules, carbon nanotubes and
graphene are oxidized to introduce carboxylic (–COOH) groups
on the sidewalls and edge-planes of carbon nanotubes and graphene
(Datsyuk et al., 2008); reduced graphene oxide already has
–COOH groups. These carboxylic groups couple with the amine
(–NH2) groups in protein or the amine terminated ends of an aptamer
via amide bond formation. Though effective, covalent modifi-
cations disrupt the in-plane sp2 bonding in the carbon lattice
pattern and introduce defects that significantly affect the desirable
electrical properties of the nanomaterials (Banerjee et al., 2005).
Non-covalent functionalization of carbon nanomaterials, on the
other hand, modifies the electrode surface in a non-destructive
manner and also enhances the solution phase stability of the nanomaterials
by preventing restacking or aggregation. This can be
achieved by either direct physical adsorption of the biomolecules
on the surface of the SWNT/graphene or using a bifunctional linker
molecule that can be grafted to the sidewalls of CNTs or planar graphene
(Chen et al., 2001). An example of such bilinker is 1-
pyrenebutanoic acid succinimidyl ester (PBASE) in which the pyrene
binds to SWNT non-covalently through p–p interaction and
the succinimidyl ester forms an amide bond with the amine group
of a biomolecule (Fig. 6). In order to prevent the non-specific binding
of the biomolecules, the uncoated areas of graphene/CNT can
be coated with a surfactant such as Tween 20 or using an inert protein
bovine serum albumin (BSA)