Biotinsilane compounds and a binding matrix containing these compounds
AssigneeBoehringer Mannheim GmbH
InventorsPeter Sluka, Hans-Georg Batz
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Abstract
The invention concerns a binding matrix containing a carrier material with an oxidic surface and a solid phase reactant covalently bound thereto via anchor groups which is capable of binding to at least one free reaction partner, which is characterized in that the solid phase reactant forms a diluted and essentially laterally homogeneous binding layer on the surface of the carrier material and that the anchor groups are silane groups and are linked to the solid phase reactant via a spacer molecule.
Claims62
Binding matrix comprising an oxidic surface, wherein the oxidic surface is at least partially covered by a diluted and essentially laterally homogeneous binding layer, said binding layer formed by covalent reaction of a silane compound with the oxidic surface, said silane compound selected from the group consisting of:
- ##STR26## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are the same or different and are substituents on the Si atom, provided that at least one of the substituents is covalently bound to the oxidic surface, and wherein said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino and halogen;
- Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms, and X.sup.1 X.sup.2 and X.sup.3 are each solid phase reactants capable of binding to an analyte in a sample, wherein said binding matrix further comprises diluent molecules which are covalently bound to the oxidic surface but are incapable of binding to the analyte.
Method for the determination of an analyte in a sample solution comprising providing a binding matrix comprising an oxidic surface wherein the oxidic surface is partially covered by a diluted and essentially laterally homogeneous binding layer, said binding layer formed by covalent reaction of a silane compound with the oxidic surface, said silane compound selected from the group consisting of:
- ##STR27## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are the same or different and are substituents on the Si atom, provided that at least one of the substituents is covalently bound to the oxidic surface, wherein said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino and halogen;
- Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms, and X.sup.1, X.sup.2 and X.sup.3 are each solid phase reactants capable of binding to an analyte in a sample, wherein said binding matrix further comprises diluent molecules which are covalently bound to the oxidic surface but incapable of binding to the analyte in a sample, and contacting said binding matrix with a solution containing said analyte, said analyte having a bioaffinity for the solid phase reactant of the binding matrix, under conditions favoring binding of said analyte to said binding matrix;
- and detecting the bound analyte.
Method for the concurrent determination of different analytes in a sample solution, comprising contacting the sample solution with at least one member selected from the group consisting of:
- (a) a compartmented binding matrix comprising a plurality of spatially separated zones, each zone comprising a binding matrix having an oxidic surface, wherein the oxidic surface is at least partially covered by a diluted and essentially laterally homogenous binding layer, said binding layer formed by covalent reaction of a silane compound with the oxidic surface, said silane compound selected from the group consisting of:
- ##STR28## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are the same or different and are substituents on the Si atom, provided that at least on of the substituents is covalently bound to the oxidic surface and said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino and halogen;
- Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms, and X.sup.1, X.sup.2 and X.sup.3 are each solid phase reactants capable of binding to an analyte in a sample, wherein said binding matrix further comprises diluent molecules which are covalently bound to the oxidic surface but are incapable of binding to any of the analytes, and wherein the spatially separated zones comprise binding matrices with different solid phase reactants, and
- (b) an analytical element comprising a compartmented binding matrix of (a), wherein the solid phase reactant is covered with one or more layers of reaction partners, said compartmented binding matrix comprising a plurality of spatially separated zones, each of which comprises a binding matrix as in (a), and wherein the individual spatially separated zones are each covered with different reaction partners;
- and
- detecting the presence of analyte in said sample solution by locally resolving measurement of individually spatially separated binding matrix zones.
Method for the concurrent determination of an analyte in different sample solutions, wherein in each determination a sample solution is contacted with individual zones of at least one member selected from the group consisting of:
- (a) a compartmented binding matrix comprising a plurality of spatially separated zones, each zone comprising a binding matrix comprising an oxidic surface, wherein the oxidic surface is at least partially covered by a diluted and essentially laterally homogenous binding layer, said binding layer formed by covalent reaction of a silane compound with the oxidic surface, said silane compound selected from the group consisting of:
- ##STR29## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are the same or different and are substituents on the Si atom, provided that at least one of the substituents is covalently bound to the oxidic surface and said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino and halogen;
- Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms, and X.sup.1, X.sup.2 and X.sup.3 are each solid phase reactants capable of binding to an analyte in a sample, wherein said binding matrix further comprises diluent molecules which are covalently bound to the oxidic surface but which are incapable of binding to an analyte, and wherein the spatially separated zones comprise binding matrices each with the same solid phase reactant, and
- (b) an analytical element comprising a compartmented binding matrix of (a), wherein the solid phase reactant is covered with one or more layers of reaction partners, and wherein the spatially separated zones are each covered with the same reaction partners, and
- detecting the presence of the analyte in each sample solution by locally resolving measurement of individually separated binding matrix zones.
A silane compound selected from the group consisting of:
- ##STR30## wherein R.sup.1, R.sup.2, R.sup.3, and R.sup.4 can be the same or different and are substituents on the Si atom, provided that at least one of the substituents R.sup.1, R.sup.2, R.sup.3 and R.sup.4, can covalently bind to an oxidic surface, wherein said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino, and halogen, Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms which is covalently bound to the Si atom by an Si--C bond, and X.sup.1, X.sup.2, X.sup.3 are selected from the group consisting of biotin, a biotin analogue, a biotin derivative, a hapten, an oligonucleotide, a polynucleotide, and a C.sub.1 -C.sub.1 alkoxy(oligo-C.sub.2 -C.sub.4 alkylene-oxide) group and are covalently bound to the spacer molecule.
Process for making an article having an oxidic surface which has decreased unspecific protein binding to oxidic surfaces comprising bringing the oxidic surface into contact with a solution containing diluent molecules comprising at least one silane compound selected from the group consisting of:
- ##STR31## wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are the same or different and are substituents on the Si atom, provided that at least one of the substituents R.sup.1, R.sup.2, R.sup.3 and R.sup.4 can covalently bind to an oxidic surface, and said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino and halogen;
- Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms which is covalently bound to the Si atom by an Si--C bond, and X.sup.1, X.sup.2 and X.sup.3 are each covalently bound to the spacer molecule, under conditions which lead to a covalent reaction of at least one of said silane compounds with the oxidic surface, and wherein X.sup.1, X.sup.2, and X.sup.3 are C.sub.1 -C.sub.4 alkoxy(oligo-C.sub.2 -C.sub.4 -alkylene-oxide) groups.
Article comprising an oxidic surface, wherein the oxidic surface is covered by a binding layer which is formed by the covalent reaction of diluent molecules comprising at least one silane compound selected from the group consisting of:
- ##STR32## wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are the same or different and are substituents on the Si atom, provided that at least one of the substituents R.sup.1, R.sup.2, R.sup.3 and R.sup.4 covalently binds to the oxidic surface, wherein said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino and halogen;
- Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms which is covalently bonded to the Si atom by an Si--C bond and X.sup.1, X.sup.2 and X.sup.3 are covalently bound to the spacer molecule, wherein X.sup.1, X.sup.2 and X.sup.3 are C.sub.1 -C.sub.4 alkoxy(oligo-C.sub.2 -C.sub.4 alkylene oxide) groups.
A process for the production of a binding matrix comprising an oxidic surface wherein the oxidic surface is at least partially covered by an essentially laterally homogeneous binding layer said process comprising contacting the oxidic surface with a solution comprising a silane compound selected from the group consisting of:
- ##STR33## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are the same or different and are substituents on the Si atom, provided that at least one of the substituents is able to covalently bind to the oxidic surfaces, wherein said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino and halogen;
- Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms, and X.sup.1, X.sup.2 and X.sup.3 are each solid phase reactants capable of binding to an analyte in a sample;
- and
- diluent molecules capable of covalently binding to the oxidic surface but incapable of binding to an analyte in a sample, wherein said silane compound is present at a concentration of from 10.sup.-4 to 10.sup.-2 % by weight, under conditions favoring covalent attachment of said silane compound and said diluent molecules to the oxidic surface.
A process for the production of a compartmented binding matrix comprising a plurality of spatially separated zones, each zone comprising a binding matrix comprising an oxidic surface, wherein the oxidic surface is at least partially covered by an essentially laterally homogeneous binding layer, said process comprising contacting each oxidic surface of the binding matrices with a solution comprising a silane compound selected from the group consisting of:
- ##STR34## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are the same or different and are substituents on the Si atom, provided that at least one of the substituents is able to covalently bind to the oxidic surfaces;
- wherein said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino and halogen;
- Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms, and X.sup.1, X.sup.2 and X.sup.3 are each solid phase reactants capable of binding to an analyte in a sample;
- and
- diluent molecules capable of covalently binding to the oxidic surface but incapable of covalently binding to an analyte in the sample, wherein the concentration of the silane compound is 10.sup.-4 to 10.sup.-2 % by weight, under conditions favoring covalent attachment of said silane compound and said diluent molecules to the oxidic surface.
A process for the production of a binding matrix comprising an oxidic surface, wherein the oxidic surface is at least partially covered by a diluted and essentially laterally homogeneous binding layer, said process comprising contacting the oxidic surface with a solution comprising a silane compound selected from the group consisting of:
- ##STR35## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are the same or different and are substituents on the Si atom, provided that at least one of the substituents is able to covalently bind to the oxidic surface wherein said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino and halogen;
- Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms, and X.sup.1, X.sup.2 and X.sup.3 are each solid phase reactants capable of binding to an analyte in a sample, said solution also comprising diluent molecules that are capable of binding to the oxidic surface but are incapable of binding to an analyte in the sample, wherein said silane compounds and said diluent molecules are present in said solution in a ratio of 1:10 to 1:2, under conditions favoring covalent attachment of said silane compounds and said diluent molecules to said oxidic surface.
A process for the production of a compartmented binding matrix comprising a plurality of spatially separated zones, each zone comprising a binding matrix comprising an oxidic surface, wherein the oxidic surface is at least partially covered by a diluted and essentially laterally homogeneous binding layer, said process comprising contacting the oxidic surface with a solution comprising a silane compound selected from the group consisting of:
- ##STR36## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are the same or different and are substituents on the Si atom, provided that at least one of the substituents is able to covalently bind to the oxidic surface;
- wherein said substituents are selected from the group consisting of C.sub.1 -C.sub.4 alkyl, C.sub.1 -C.sub.4 alkoxy, hydroxy, amino and halogen;
- Sp is a flexible, linear spacer molecule with a chain length of 2 to 50 atoms, and X.sup.1, X.sup.2 and X.sup.3 are each solid phase reactants capable of binding to an analyte in a sample, said solution also comprising diluent molecules that are capable of covalently binding to the oxidic surface but are incapable of binding to an analyte in the sample, wherein said silane compounds and said diluent molecules are present in said solution in a ratio of 1:10 to 1:2, under conditions favoring covalent attachment of said silane compounds and said diluent molecules to said oxidic surfaces.
Dependencies are inferred from the claim text.