The thermal degradation related to stability in long-term storage of a carbon nanotube-based biosensor has been investigated. The effect of storage temperature on detachment and denaturation of glucose oxidase (GOx) biocatalyst has been proved. The carbon nanotubes (CNTs) coated with polyethyleneimine (PEI) as entrapping polymer to attract more GOx to form a durable and layered CNT/PEI/GOx structure is used for long-term storage to minimize GOx detachment from the structure and minimize the possibility of enzyme and protein denaturation. After 120 days, the glucose response of the CNT/PEI/GOx biosensor stored under 4°C is preserved up to 66.7% of its initial value, while under a 25 °C storage the response is maintained up to 41.7%. The enzyme coverage activity of CNT/PEI/GOx stored at 4 °C and 25 °C has decreased by 31.1% and 51.4%, respectively. Denaturation and detachment of GOx are the common causes of thermal degradation in biosensors under improper storage temperatures, but the presence of PEI in the structure can slow-down these phenomena. Moreover, the electrons transfer constant of CNT/PEI/GOx biocatalyst stored at 4 °C and 25 °C were 7.5 ± 0.5 s−1
and 6.6 ± 0.3 s−1
, respectively, indicating that also electrons mobility is damaged by detachment and denaturation of enzyme protein and the detection of glucose from the glucose oxidation reaction (GOR) is compromised.
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