A graphene oxide based fluorescence resonance energy transfer (FRET) biosensor for ultrasensitive detection of botulinum neurotoxin A (BoNT/A) enzymatic activity

Jingyu Shi, Jiubiao Guo, Gongxun Bai, Chunyu Chan, Xuan Liu, Weiwei Ye, Jianhua Hao, Sheng Chen, Mo Yang

Research output: Journal article publicationJournal articleAcademic researchpeer-review

43 Citations (Scopus)

Abstract

Botulinum neurotoxins (BoNTs) are among the most potent toxic bacterial proteins for humans, which make them potential agents for bioterrorism. Therefore, an ultrasensitive detection of BoNTs and their active states is in great need as field-deployable systems for anti-terrorism applications. We report the construction of a novel graphene oxide (GO)-peptide based fluorescence resonance energy transfer (FRET) biosensor for ultrasensitive detection of the BoNT serotype A light chain (BoNT-LcA) protease activity. A green fluorescence protein (GFP) modified SNAP-25 peptide substrate (SNAP-25-GFP) was optimally designed and synthesized with the centralized recognition/cleavage sites. This FRET platform was constructed by covalent immobilization of peptide substrate on GO with BSA passivation which have advantages of low non-specific adsorption and high stability in protein abundant solution. BoNT-LcA can specifically cleave SNAP-25-GFP substrate covalently immobilized on GO to release the fragment with GFP. Based on fluorescence signal recovery measurement, the target BoNT-LcA was detected sensitively and selectively with the linear detection range from 1. fg/mL to 1. pg/mL. The limit of detection (LOD) for BoNT-LcA is around 1. fg/mL.
Original languageEnglish
Pages (from-to)238-244
Number of pages7
JournalBiosensors and Bioelectronics
Volume65
DOIs
Publication statusPublished - 5 Mar 2015

Keywords

  • Botulinum neurotoxins (BoNTs)
  • Graphene oxide (GO)
  • Green fluorescence protein (GFP)
  • Optical detection
  • SNAP-25

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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