Biosensor involving the use of optically sensitive moieties

A method of fabricating a biosensor in which there is at least one discrete test and at least one discrete control zone, the method comprising the following steps:

• (i) providing a conductive substrate;
• (ii) forming a membrane including membrane spanning lipids and ion channels comprising first and second half membrane spanning monomers, the membrane being attached to the conductive substrate in amanner such that a functioning reservoir exists between the membrane and the conductive substrate;
• (iii) providing on the ion channels and membrane spanning lipids a photoactivatable group which when illuminated will bind a receptor;
• (iv) illuminating discrete areas of the membrane and linking a ligand reactive with an analyte of interest to the ion-channel and membrane spanning lipid components of the tethered membrane via photoactivated group to form test areas;
• (v) removing unbound ligand;
• and
• (vi) linking a control ligand to the remainder of the ion-channels and membrane spanning lipid components of the tethered membrane to form control areas.

A method of producing an improved biosensor, the method comprising the following steps:

• (i) providing a conductive substrate;
• (ii) forming a membrane including membrane spanning lipids and ion channels comprising first and second half membrane spanning monomers, the membrane being attached to the conductive substrate such that a functioning reservoir exists between the membrane and the conductive substrate;
• (iii) providing on the half membrane spanning monomer remote from the conductive substrate a photoswitchable binder to streptavidin and providing biotin on the membrane spanning lipid;
• (iv) adding streptavidin;
• (v) triggering a light source and rinsing;
• (vi) triogering the light source off and adding streptavidin;
• (vii) optionally repeating steps (v) and (vi);
• and
• (viii) adding ligands specfic to an analyte to the ion channels and membrane spanning lipids.

An improved biosensor, the biosensor comprising a membrane and an electrode having a conductive substrate, the membrane including membrane spanning lipids and ion channels comprising first and second half membrane spanning monomers, the membrane being attached to the conductive substrate such that a functioning reservoir exists between the membrane and the conductive substrate, and ligands specific for an analyte attached to the ion channels and membrane spanning lipids, the improvement comprising providing on at least one of the first and second half membrane spanning monomers or membrane spanning lipids a photoswitchable group derived from a compound in accordance with Formula 1:

• 4
• wherein R1 represents 0 to about 3 groups where each is independently H or saturated or unsaturated, substituted or unsubstituted C1-C10 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon;
• R2 represents 0 to about 3 groups where each is independently hydrogen or satmuated or unsaturated, substituted or unsubstituted C1-10 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon;
• Y represents H, saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon, COR6, CONR7R8, COOR14, S(O)nR15 where n is 0, 1 or 2, R6, R7, R8, R14 and R15 are each independently represent H, saturated or unsaturated, substituted or unsubstituted C1-10 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4, hydrocarbon or aryl;
• R9 is —C(O)X where X represents H, saturated or unsaturated, substituted or unsubstituted C1-10 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon, or OH, or OR10 in which R10 is alkyl, or NR11R12 in which R11 and R12 are H, alkyl or taken together with N form a rincg, or arl or R9 together with R1 form a substituted or unsubstituted 5-6 member cyclic or heterocyclic ring;
• Z represents O or NR13 R13 is H, saturated or unsaturated, substituted or unsubstituted C1-10 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon or aryl, the compound including a functional group at Y or R9 such that the compound can be linked to the the at least one of the first and second half membrane spanning monomers.

A compound in accordance with Formula 1:

• 6
• wherein R1 represents 0 to about 3 groups where each is independently H or saturated or unsuaturated, substituted or usubstituted C1-10 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon;
• R2 represents 0 to about 3 groups where each is independently hydrogen or saturated or unsaturated, substituted or unsubstituted C1-10 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon;
• Y represents H, saturated or unsaturated, substituted or unsubstituted C1-10 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon, COR6, CONR7R8, COOR14, S(O)nR15 where n is 0, 1, or 2, R6, R7, R14 and R15 are each independently represent H, saturated or unsaturated, substituted or unsubstitusted C1-10 hydrocarbon, preferably, saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon or aryl;
• R9 is —C(O)X where X represents H, saturated or unsaturated, substituted or unsubstituted C1-10 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon, or OH, or OR10 in which R10 is alkyl, or NR11R12 in which R11 and R12 are H, alkyl or taken together with N form a ring, or aryl or R9 together with R1 form a substituted or usubstituted 5-6 member cyclic or heterocyclic ring;
• Z represents O or NR13 R13 is H, saturated or unsaturated, substituted or unsubstituted C1-10 hydrocarbon, preferably saturated or unsaturated, substituted or unsubstituted C1-4 hydrocarbon or aryl, with the proviso that when R is —C(O)X and X is H, at least one of R1, R2, R3 or Y is other than H.