PQQ-Dependent Glucose Dehydrogenase as Potential Enzymatic Label in Amperometric Avidin-Biotin Systems

In the current work, PQQ‐dependent glucose dehydrogenase (s‐PQQ‐GDH) has been investigated as a potential biotin labeling agent. The conjugation of s‐PQQ‐GDH with biotin was achieved according to newly proposed technique using a commercially available biotin pentafluorophenyl ester. The residual act...

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Bibliographic Details
Published in:Electroanalysis (New York, N.Y.) N.Y.), 2007-01, Vol.19 (2-3), p.280-285
Main Authors: Razumiene, J., Vilkanauskyte, A., Gureviciene, V., Bachmatova, I., Marcinkeviciene, L., Laurinavicius, V., Meskys, R.
Format: Article
Language:English
Subjects:
Amperometric analysis
Carbon and Gold electrodes
s-PQQ-GDH
s-PQQ-GDH labeled biotin
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Summary:In the current work, PQQ‐dependent glucose dehydrogenase (s‐PQQ‐GDH) has been investigated as a potential biotin labeling agent. The conjugation of s‐PQQ‐GDH with biotin was achieved according to newly proposed technique using a commercially available biotin pentafluorophenyl ester. The residual activity of conjugate was about 90% against 100% of the native activity. The suitability of the conjugate for amperometric immunoanalysis was tested using carbon (CE) and gold (Au) electrodes. An efficient electron transfer from the active centre of s‐PQQ‐GDH to the CE or Au electrode surface mediated by phenazine methasulfate (PMS) allowed application of the s‐PQQ‐GDH as a biotin label in amperometric analysis based on interaction with neutravidin. The optimization of the amperometric analysis steps such as coating, blocking and binding were performed.
ISSN:1040-0397
1521-4109
DOI:10.1002/elan.200603715