Selection of affinity peptides for interference-free detection of cholera toxin.

Autor: Lim JM; Department of Pharmaceutical Engineering, Daegu Haany University, Gyeongsan 38610, Republic of Korea., Heo NS; Department of Biological Engineering, Inha University, 100 Inha-ro, Nam-gu, Incheon 22212, Republic of Korea., Oh SY; Department of Biological Engineering, Inha University, 100 Inha-ro, Nam-gu, Incheon 22212, Republic of Korea., Ryu MY; Department of Pharmaceutical Engineering, Daegu Haany University, Gyeongsan 38610, Republic of Korea., Seo JH; School of Chemical Engineering, Yeungnam University, Gyeongsan 38541, Republic of Korea., Park TJ; Department of Chemistry, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul 06974, Republic of Korea., Huh YS; Department of Biological Engineering, Inha University, 100 Inha-ro, Nam-gu, Incheon 22212, Republic of Korea. Electronic address: yunsuk.huh@inha.ac.kr., Park JP; Department of Pharmaceutical Engineering, Daegu Haany University, Gyeongsan 38610, Republic of Korea. Electronic address: jppark@dhu.ac.kr.
Jazyk: angličtina
Zdroj: Biosensors & bioelectronics [Biosens Bioelectron] 2018 Jan 15; Vol. 99, pp. 289-295. Date of Electronic Publication: 2017 Jul 31.
DOI: 10.1016/j.bios.2017.07.075
Abstrakt: Cholera toxin is a major virulent agent of Vibrio cholerae, and it can rapidly lead to severe dehydration, shock, causing death within hours without appropriate clinical treatments. In this study, we present a method wherein unique and short peptides that bind to cholera toxin subunit B (CTX-B) were selected through M13 phage display. Biopanning over recombinant CTX-B led to rapid screening of a unique peptide with an amino acid sequence of VQCRLGPPWCAK, and the phage-displayed peptides analyzed using ELISA, were found to show specific affinities towards CTX-B. To address the use of affinity peptides in development of the biosensor, sequences of newly selected peptides were modified and chemically synthesized to create a series of affinity peptides. Performance of the biosensor was studied using plasmonic-based optical techniques: localized surface plasmon resonance (LSPR) and surface-enhanced Raman scattering (SERS). The limit of detection (LOD) obtained by LSPR with 3σ-rule was 1.89ng/mL, while SERS had a LOD of 3.51pg/mL. In both cases, the sensitivity was much higher than the previously reported values, and our sensor system was specific towards actual CTX-B secreted from V. cholera, but not for CTX-AB 5 .
(Copyright © 2017 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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