Journal
BIOSENSORS & BIOELECTRONICS
Volume 38, Issue 1, Pages 233-238Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2012.05.036
Keywords
Aptamer; Nampt (PBEF/visfatin); SELEX; Capacitive biosensor
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Funding
- Ministry for Health, Welfare and Family Affairs, Republic of Korea [A081023]
- National Research Foundation (NRF) of Korea
- Korea government (Ministry of Education, Science and Technology [MEST]) [2010-00579-KORANET]
- Scientific and Technological Research Council of Turkey (TUBITAK) [110E287]
- Korea Health Promotion Institute [A081023] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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A single-stranded DNA (ssDNA) aptamer was successfully developed to specifically bind to nicotinamide phosphoribosyl transferase (Nampt) through systematic evolution of ligands by exponential enrichment (SELEX) and successfully implemented in a gold-interdigitated (GID) capacitor-based biosensor. Surface plasmon resonance (SPR) analysis of the aptamer revealed high specificity and affinity (K-d = 72.52 nM). Changes in surface capacitance/charge distribution or dielectric properties in the response of the GID capacitor surface covalently coupled to the aptamers in response to changes in applied AC frequency were measured as a sensing signal based on a specific interaction between the aptamers and Nampt. The limit of detection for Nampt was 1 ng/ml with a dynamic serum detection range of up to 50 ng/ml; this range includes the clinical requirement for both normal Nampt level, which is 15.8 ng/ml, and Nampt level in type 2 diabetes mellitus (T2DM) patients, which is 31.9 ng/ml. Additionally, the binding kinetics of aptamer-Nampt interactions on the capacitor surface showed that strong binding occurred with increasing frequency (range, 700 MHz-1 GHz) and that the dissociation constant of the aptamer under the applied frequency was improved 120-240 times (K-d = 0.3-0.6 nM) independent on frequency. This assay system is an alternative approach for clinical detection of Nampt with improved specificity and affinity. (C) 2012 Elsevier B.V. All rights reserved.
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