Immunoreaction measurement method
View Patent ↗The present invention relates to an immunoreaction measuring method for measuring an antigen or antibody contained as a subject substance in a sample, wherein the sample was mixed with at least one compound selected from the group consisting of dicarboxylic acids having a hydroxyl group, dicarboxylic acids having a double bond, straight-chain dicarboxylic acids expressed by the chemical formula (1): HOOC(CH 2 ) n COOH (n is an integer), and the salts of these dicarboxylic acids, and an antibody or antigen as a specifically binding substance capable of specifically binding to the subject substance, to obtain an acidic reaction solution, and then an antigen-antibody complex, generated by an antigen-antibody reaction of the subject substance with the specifically binding substance in the reaction solution was detected. This makes it possible to improve a measurement value and to relax a limitation of a measurement range due to a zone phenomenon that occurs in an antigen-excess region.
1. An immunoreaction measurement method for measuring an antigen or antibody contained as a subject substance in a sample, comprising the steps of:
(A) mixing said sample; at least one compound selected from the group consisting of tartaric acid, itaconic acid, straight-chain dicarboxylic acids expressed by the chemical formula (1): HOOC(CH 2 ) n COOH (n is an integer from 1 to 7), and the salts of these dicarboxylic acids; and an antibody or antigen as a specifically binding substance capable of specifically binding to said subject substance, to obtain an acidic reaction solution; and
(B) detecting in said reaction solution an antigen-antibody complex generated by an antigen-antibody reaction of said subject substance and said specifically binding substance,
wherein the pH of said reaction solution is set to 5.0 to 6.0 when said compound comprises at least one selected from the group consisting of said straight-chain dicarboxylic acids and salts of these dicarboxylic acids, and
the pH of said reaction solution is set to 4.5 to 5.0 when said compound comprises at least one selected from the group consisting of said tartaric acid, said itaconic acid and salts of these dicarboxylic acids.
2. The immunoreaction measurement method in accordance with claim 1 , wherein said reaction solution is further mixed with a buffer in said step (A).
3. The immunoreaction measurement method in accordance with claim 1 , wherein, in said reaction solution, the concentration of said at least one compound selected from the group consisting of tartaric acid, itaconic acid straight-chain dicarboxylic acids expressed by the chemical formula (1): HOOC(CH 2 ) n COOH (n is an integer from 1 to 7), and the salts of these dicarboxylic acids, is set to not higher than 0.1 M.
4. The immunoreaction measurement method in accordance with claim 1 , wherein, in said reaction solution, the concentration of said at least one compound selected from the group consisting of tartaric acid, itaconic acid, straight-chain dicarboxylic acids expressed by the chemical formula (1): HOOC(CH 2 ) n COOH (n is an integer from 1 to 7), and the salts of these dicarboxylic acids, is set to the range of 0.01 M to 0.1 M.
5. The immunoreaction measurement method in accordance with claim 1 , wherein, in said reaction solution, the concentration of said at least one compound selected from the group consisting of tartaric acid, itaconic acid, straight-chain dicarboxylic acids expressed by the chemical formula (1): HOOC(CH 2 ) n COOH (n is an integer from 1 to 7), and the salts of these dicarboxylic acids, is set to the range of 0.01 M to 0.05 M.
6. The immunoreaction measurement method in accordance with claim 1 , wherein said reaction solution contains polyethylene glycol in an amount of not smaller than 2 wt % and not larger than 6 wt %.
7. The immunoreaction measurement method in accordance with claim 1 , wherein said antigen-antibody complex is an agglutination complex.
8. The immunoreaction measurement method in accordance with claim 7 , wherein said agglutination complex is detected by measuring optical variations attributed to said agglutination complex.
9. The immunoreaction measurement method in accordance with claim 8 , wherein said optical variations are variations in intensity of scattered light.
10. The immunoreaction measurement method in accordance with claim 1 , wherein said specifically binding substance is an antibody comprising a monoclonal antibody.
11. The immunoreaction measurement method in accordance with claim 1 , wherein said specifically binding substance includes at least two sort or more of monoclonal antibodies.
12. The immunoreaction measurement method in accordance with claim 1 , wherein said antigen is human albumin.