TY - JOUR
T1 - Glutamate receptor incorporated in a mixed hybrid bilayer lipid membrane array, as a sensing element of a biosensor working under flowing conditions
AU - Favero, Gabriele
AU - Campanella, Luigi
AU - Cavallo, Stefano
AU - D'Annibale, Andrea
AU - Perrella, Maurizio
AU - Mattei, Elisabetta
AU - Ferri, Tommaso
PY - 2005/6/8
Y1 - 2005/6/8
N2 - The realization of a reliable receptor biosensor requires stable, long-lasting, reconstituted biomembranes able to supply a suitable biomimetic environment where the receptor can properly work after incorporation. To this end, we developed a new method for preparing stable biological membranes that couple the biomimetic properties of BLMs (bilayer lipid membranes) with the high stability of HBMs (hybrid bilayer membranes); this gives rise to an innovative assembly, named MHBLM (mixed hybrid bilayer lipid membrane). The present work deals with the characterization of biosensors achieved by embedding an ionotropic glutamate receptor (GluR) on MHBLM. Thanks to signal (transmembrane current) amplification, which is typical of natural receptors, the biosensor here produced detects glutamate at a level of nmol L-1. The transmembrane current changes linearly vs glutamate up to 100 nmol L-1, while the limit of detection is 1 nmol L-1. In addition, the biosensor response can be modulated both by receptor agonists (glycine) and antagonists (Mg2-) as well, and by exploiting the biosensor response, the distribution of different kinds of ionotropic GluR present in the purified sample, and embedded in MHBLM, was also evaluated. Finally, one of the most important aspects of this investigation is represented by the high stability of the biomimetic system, which allows the use of biosensor under flowing conditions, where the solutions flow on both biomembrane faces.
AB - The realization of a reliable receptor biosensor requires stable, long-lasting, reconstituted biomembranes able to supply a suitable biomimetic environment where the receptor can properly work after incorporation. To this end, we developed a new method for preparing stable biological membranes that couple the biomimetic properties of BLMs (bilayer lipid membranes) with the high stability of HBMs (hybrid bilayer membranes); this gives rise to an innovative assembly, named MHBLM (mixed hybrid bilayer lipid membrane). The present work deals with the characterization of biosensors achieved by embedding an ionotropic glutamate receptor (GluR) on MHBLM. Thanks to signal (transmembrane current) amplification, which is typical of natural receptors, the biosensor here produced detects glutamate at a level of nmol L-1. The transmembrane current changes linearly vs glutamate up to 100 nmol L-1, while the limit of detection is 1 nmol L-1. In addition, the biosensor response can be modulated both by receptor agonists (glycine) and antagonists (Mg2-) as well, and by exploiting the biosensor response, the distribution of different kinds of ionotropic GluR present in the purified sample, and embedded in MHBLM, was also evaluated. Finally, one of the most important aspects of this investigation is represented by the high stability of the biomimetic system, which allows the use of biosensor under flowing conditions, where the solutions flow on both biomembrane faces.
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U2 - 10.1021/ja042904g
DO - 10.1021/ja042904g
M3 - Article
C2 - 15926837
AN - SCOPUS:20444407699
VL - 127
SP - 8103
EP - 8111
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
SN - 0002-7863
IS - 22
ER -