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This paper reports a study of the factors affecting the analytical performance of gold and glassy carbon electrodes modified with the tripeptide Gly-Gly-His for the detection of copper ions. Gly-Gly-His is attached to a glassy carbon (GC) surface modified with 4-carboxyphenyl moieties or a gold surface modified with 3-mercaptopropionic acid by the reaction of the N-terminal amine group of the peptide with the carboxylic acid groups of the monolayer via carbodiimide activation. X-ray photoelectron spectroscopy was used to characterize the steps in the biosensor fabrication. It was found that the analytical performance of a sensor prepared with Gly-Gly-His on a GC electrode was similar to that on a gold electrode under the same conditions. The performance was greatly enhanced at higher temperature, no added salt during copper accumulation and longer accumulation time within a pH range of 7-9. Interference studies and investigations of stability of the Gly-Gly-His sensor are reported. Analysis of natural water samples show that the sensors measure only copper ions that can complex at the sensor surface. Strongly complexed copper in natural water is not measured. Despite greater stability of diazonium salt derived monolayers on carbon surfaces compared with alkanethiols self-assembled monolayers on gold, the stability of the sensors was essentially the same regardless of the modification procedure.