Specific and reversible immobilization of proteins tagged to the affinity polypeptide C-LytA on functionalized graphite electrodes

Please use this identifier to cite or link to this item: http://hdl.handle.net/10045/35464
Información del item - Informació de l'item - Item information
Title: Specific and reversible immobilization of proteins tagged to the affinity polypeptide C-LytA on functionalized graphite electrodes
Authors: Bello Gil, Daniel | Maestro, Beatriz | Fonseca, Jennifer | Feliu, Juan M. | Climent, Victor | Sanz, Jesús M.
Research Group/s: Electroquímica de Superficies
Center, Department or Service: Universidad de Alicante. Departamento de Química Física | Universidad de Alicante. Instituto Universitario de Electroquímica
Keywords: Specific and reversible immobilization | Proteins | C-LytA | Graphite electrodes
Knowledge Area: Química Física
Issue Date: 31-Jan-2014
Publisher: Public Library of Science (PLoS)
Citation: Bello-Gil D, Maestro B, Fonseca J, Feliu JM, Climent V, et al. (2014) Specific and Reversible Immobilization of Proteins Tagged to the Affinity Polypeptide C-LytA on Functionalized Graphite Electrodes. PLoS ONE 9(1): e87995. doi:10.1371/journal.pone.0087995
Abstract: We have developed a general method for the specific and reversible immobilization of proteins fused to the choline-binding module C-LytA on functionalized graphite electrodes. Graphite electrode surfaces were modified by diazonium chemistry to introduce carboxylic groups that were subsequently used to anchor mixed self-assembled monolayers consisting of N,N-diethylethylenediamine groups, acting as choline analogs, and ethanolamine groups as spacers. The ability of the prepared electrodes to specifically bind C-LytA-tagged recombinant proteins was tested with a C-LytA-β-galactosidase fusion protein. The binding, activity and stability of the immobilized protein was evaluated by electrochemically monitoring the formation of an electroactive product in the enzymatic hydrolysis of the synthetic substrate 4-aminophenyl β-D-galactopyranoside. The hybrid protein was immobilized in an specific and reversible way, while retaining the catalytic activity. Moreover, these functionalized electrodes were shown to be highly stable and reusable. The method developed here can be envisaged as a general, immobilization procedure on the protein biosensor field.
Sponsor: This work was supported by grants BFU2010–17824, CTQ2008-04492-E and CTQ2010-18570 (Spanish Ministerio de Economia y Competitividad) and FPA-2010-055 (Valentian Government, Spain).
URI: http://hdl.handle.net/10045/35464
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0087995
Language: eng
Type: info:eu-repo/semantics/article
Rights: © 2014 Bello-Gil et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Peer Review: si
Publisher version: http://dx.doi.org/10.1371/journal.pone.0087995
Appears in Collections:INV - EQSUP - Artículos de Revistas

Files in This Item:
Files in This Item:
File Description SizeFormat 
Thumbnail2014_Bello-Gil_etal_PLoS-ONE.pdf601,39 kBAdobe PDFOpen Preview


This item is licensed under a Creative Commons License Creative Commons