RT Journal Article
T1 Specific and reversible immobilization of proteins tagged to the affinity polypeptide C-LytA on functionalized graphite electrodes
A1 Bello-Gil, Daniel
A1 Maestro García-Donas, María Beatriz
A1 Fonseca, Jennifer
A1 Feliu, Juan
A1 Climent, Victor
A1 Sanz, Jesús
AB 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.
PB Public Library of Science
SN 1932-6203
YR 2014
FD 2014
LK https://hdl.handle.net/20.500.14352/93157.2
UL https://hdl.handle.net/20.500.14352/93157.2
LA eng
NO Bello-Gil D, Maestro B, Fonseca J, Feliu JM, Climent V, Sanz JM (2014) Specific and Reversible Immobilization of Proteins Tagged to the Affinity Polypeptide C-LytA on Functionalized Graphite Electrodes. PLoS ONE 9(1): e87995. https://doi.org/10.1371/journal.pone.0087995
NO Ministerio de Economía, Industria y Competitividad (España)
NO Generalitat Valenciana
DS Docta Complutense
RD 8 abr 2025