Chemistry: electrical and wave energy – Apparatus – Electrolytic
Patent
1993-09-21
1995-03-28
Niebling, John
Chemistry: electrical and wave energy
Apparatus
Electrolytic
204403, 20415312, 2041531, 435817, 435291, G01N 2726
Patent
active
054013780
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to electrode membrane combinations for use in biosensors. In addition, the present invention relates to methods for the production of such electrode membrane combinations and the use of biosensors incorporating such electrode membrane combinations in the detection of analytes. The present invention also relates to novel compounds used in the electrode membrane combinations.
BACKGROUND OF THE INVENTION
Biosensors comprising membranes incorporating gated ion channels have been disclosed in International Patent Application Nos. WO89/01159 and WO90/08783. As is disclosed in these applications, suitably modified receptor molecules may be caused to co-disperse with amphiphilic molecules and produce membranes with altered surface binding properties, which are useful in the production of biosensor receptor surfaces of high-binding ability and high-binding specificities. It is also disclosed that ion channels such as polypeptide ionophores may be co-dispersed with amphiphilic molecules, thereby forming membranes with altered properties in relation to the permeability of ions. There is also disclosure in these applications of various methods of gating these ion channels such that in response to the binding of an analyte the conductivity of the membrane is altered.
DESCRIPTION OF THE INVENTION
The present inventors have now determined that the stability and ion flux properties of such membranes formed on solid substrates can be greatly improved by chemisorbing or covalently attaching an array of amphiphilic compounds onto an electrode surface such that a space is formed between the electrode and the membrane.
Prior art has taught the use of such concepts as hydrogels or porous silicon as reservoirs onto which bilayer membranes may be formed. These processes are difficult to reproduce and to date no reliably and reproducibly functioning examples have been cited in the art. The present invention differs from the prior art in that it consists of a single molecule that contains within it membrane compatible hydrophobic groups, ion compatible hydrophilic groups and groups capable of chemisorbing or covalently attaching onto an electrode surface linked in such a way so as to produce functioning amphiphilic membrane/reservoir/electrode assemblies. This invention also allows for a much simpler manufacturing process yielding far more controllable, reproducible, and stable bilayer and monolayer membranes that allow the incorporation of functioning ionophores or ion channels.
Accordingly, in a first aspect, the present invention consists in an electrode membrane combination comprising an electrode and a membrane comprising a closely packed array of amphiphilic molecules and a plurality of ionophores the membrane being connected to the electrode by means of a linker molecule(s) in a manner such that a hydrophilic space is provided between the membrane and the electrode, the space being sufficient to allow the flux of ions through the ionophores, the linker molecule comprising within the same molecule a hydrophobic region which constitutes at least a proportion of the amphiphilic molecules making up the membrane or is attached to or is embedded in the membrane, an attachment region attached to the electrode and a hydrophilic region intermediate said hydrophobic and attachment regions and spanning said space between the membrane and the electrode.
In a preferred embodiment of the invention the distance between the membrane and the electrode is in the range 10 to 1000 .ANG. and more preferably is in the range 15-100 .ANG..
In a further preferred embodiment of the invention the hydrophobic region of the linker molecule constitutes at least a proportion of the amphiphilic molecules making up the membrane.
In a preferred form of the invention when the membrane is a monolayer all the amphiphilic molecules are the hydrophobic region of the linker molecules, and in the situation where the membrane is a bilayer all the amphiphilic molecules making up the layer adjacent the electr
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Cornell Bruce A.
King Lionel G.
Pace Ronald J.
Raguse Burkhard
Australian Membrane & Biotechnology Research Institute
Bell Bruce F.
Niebling John
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