Surface of base material being inhibited in non-specific adsorption
A substrate surface on which either a substance to detect analyte or an analyte per se is immobilized, which surface is formed by a treatment of substrate surface with a liquid which contains uncrosslinked polymer based on polyethylene glycol chain segment, said treatment conducted either simultaneously with the immobilization of said substance or analyte or after said substance or analyte has been immobilized on said surface. The non-specific adsorption of impurity protein or the like which is co-existent in sample for assay is significantly restrained.
1. A substrate surface on which either a substance to detect analyte or an analyte per se is immobilized,
wherein the substrate surface is formed by the following steps A) to C) performed in the following order:
A) providing a magnetic particle surface having a carboxylic acid group or a magnetic particle surface having a surface tosyl group;
B) immobilizing the substance to detect the analyte or the analyte per se on the magnetic particle surface by treatment of the magnetic particle surface with an aqueous solution containing the substance to detect analyte or the analyte per se, and
C) immobilizing uncrosslinked polymer comprising polyethylene glycol chain segment of formula (Ia) on the magnetic particle surface by treatment of the magnetic particle surface after step b) with a liquid which contains the uncrosslinked polymer comprising the polyethylene glycol chain segment of formula (Ia):
R 1 -L 1 -(CH 2 CH 2 O) n -L 2 -NH—(CH 2 CH 2 NH) m —R 2 (Ia)
wherein:
R 1 denotes hydrogen atom, methyl, formyl which is protected, amino which is protected, corboxy which is protected or hydroxyl which is protected;
R 2 denotes hydrogen atom;
L 1 denotes —(CH 2 ) p —O—, —(CH 2 ) q —COO—,
—(CH 2 ) r —S—;
L 2 denotes —(CH 2 ) k — or
k denotes 2 ;
m denotes an integer of 5 to 10,000;
n denotes an integer of 2 to 20,000; and
p, q and r independently denote and integer of 0 and 8;
wherein said treatment with the liquid containing the uncrosslinked polymer forms a covalent bond immobilizing said polymer onto the surface carboxyl group or surface tosyl group via the amino group of the uncrosslinked polymer.
2. A substrate surface of claim 1 wherein said substance to detect analyte and said analyte form a specific bonding pair.
3. A substrate surface of claim 2 wherein said substances which form a specific bonding pair are selected from the group consisting of antigen, hapten, antibody, oligonucleotide, enzyme, substrate of enzyme, sugar, lectin, hormone, receptor protein, avidin and biotin.
4. A method for producing the substrate surface of claim 1 , which comprises the following steps A) to C) performed in the following order:
A) obtaining a substrate surface which is a magnetic particle surface having a carboxylic acid group or a magnetic particle surface having a surface tosyl group;
B) bringing an aqueous solution of the substance to detect analyte or the analyte per se which has been so modified as to be immobilizable on said substrate surface into contact with said substrate surface, under a condition under which said substance to detect analyte or said analyte per se are immobilizable on said substrate surface (A; and
C) immobilizing the uncrosslinked polymer comprising polyethylene glycol chain segment of the formula (Ia) on the magnetic particle surface by treatment of the magnetic particle surface after step (B) with a liquid which contains the uncrosslinked polymer comprising the polyethylene glycol chain segment of the formula (Ia);
wherein said treatment with the liquid containing the uncrosslinked polymer forms a covalent bond immobilizing said polymer onto the surface carboxyl group or surface tosyl group via the amino group of the uncrosslinked polymer.
5. A biosensor which is equipped with a substrate surface of any one of claims 1 to 3 .
6. A substrate surface on which either a substance to detect analyte or an analyte per se is immobilized,
wherein the substrate surface is formed by the following steps A) to C) performed in the following order:
A) providing a magnetic particle surface having a carboxylic acid group or a magnetic particle surface having a surface tosyl group;
B) immobilizing the substance to detect the analyte or the analyte per se on the magnetic particle surface by treatment of the magnetic particle surface with an aqueous solution containing the substance to detect analyte or the analyte per se, and
C) immobilizing uncrosslinked polymer comprising polyethylene glycol chain segment of formula (Ia) on the magnetic particle surface by treatment of the magnetic particle surface after step b) with a liquid which contains the uncrosslinked polymer comprising the polyethylene glycol chain segment of formula (Ia):
R 1 -L 1 -(CH 2 CH 2 O) n -L 2 -NH—(CH 2 CH 2 NH) m —R 2 (Ia)
wherein:
R 1 denotes hydrogen atom, methyl, formyl which is protected, amino which is protected, corboxy which is protected or hydroxyl which is protected;
R 2 denotes hydrogen atom;
L 1 denotes —(CH 2 ) p —O—, —(CH 2 ) q —COO—,
or —(CH 2 ) r —S—;
L 2 denotes —(CH 2 ) k — or
k denotes 2 ;
m denotes an integer of 5 to 10,000;
n denotes an integer of 2 to 20,000; and
p, q and r independently denote and integer of 0 and 8;
wherein said treatment with the liquid containing the uncrosslinked polymer forms a covalent bond immobilizing said polymer onto the surface carboxyl group or surface tosyl group via the amino group of the uncrosslinked polymer.