The reaction of N-hydroxysuccinimide (NHS)
esters with freshly polished glassy carbon surfaces offers
a facile and versatile method of derivatization. Surface
concentrations larger than 10-10
mol/cm2 can thus
be easily achieved. They can be further increased when polishing
is carried out in the presence of ammonia,
which also improves their reproducibility. It is shown that the
derivatization results from the formation
of a covalent peptide linkage by reaction of the NHS ester with
superficial amino groups on the glassy
carbon surface. The peptide linkage is remarkably stable in time
and can only be hydrolyzed in very strong
basic media. 9-fluorenylmethoxycarbonyl chloride protection,
followed by preparation of the NHS ester,
by surface derivatization and by mild deprotection allows the grafting
of a molecule that contains an amino
group located remotely from the electrode surface, thus opening a route
to the attachment of a large variety
of biomolecules, for which NHS esters are available, in a position
where their degradation should be
avoided or minimized.