Method for the quantitative determination of antigens and antibodies by ratio of absorbances at different wavelengths
First Claim
1. A method for determining the concentration of an antigen in a sample, comprising:
- (a) adding the sample to a test dispersion of a finely divided carrier having an antibody affixed thereto in a dispersing medium in which the carrier is insoluble, to produce an antigen-antibody reaction therebetween;
(b) reacting the sample antigen with the carrier antibody for a predetermined reaction period;
(c) measuring the absorbance (A) of the test dispersion after the reaction period at a first wavelength (λ
1) to determine a first absorbance value (Aλ
1), and measuring the absorbance (A) of the test dispersion after the reaction period at a second wavelength (λ
2) to determine a second absorbance value Aλ
2);
(d) determining the absorbance ratio (Aλ
1 /Aλ
2) of the first absorbance value (Aλ
1) and the second absorbance value (Aλ
2) of the test dispersion; and
(e) determining the concentration of the sample antigen by comparing the absorbance ratio of the test dispersion (Aλ
1 /Aλ
2) to reference absorbance ratios at said wavelengths of reference dispersions of said carrier having said antibody affixed thereto and reacted with pre-selected concentrations of said antigen.
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Accused Products
Abstract
A method for the quantitative determination of antigen (or antibody) which comprises adding a sample containing an antigen (or antibody) to a dispersion of an insoluble carrier of fine particle size with an antibody (or antigen) fixed thereto to effect an antigen-antibody reaction, measuring absorbance of the reaction mixture, Aλ1 and Aλ2, at two different wavelengths, λ1 and λ2, and calculating the concentration of said antigen (or antibody) in the sample from the absorbance ratio Aλ1 /Aλ2. This method is suitable for the measurment of the amount of an antigen or antibody in samples taken from living bodies using a simple procedure which has high sensitivity.
25 Citations
16 Claims
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1. A method for determining the concentration of an antigen in a sample, comprising:
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(a) adding the sample to a test dispersion of a finely divided carrier having an antibody affixed thereto in a dispersing medium in which the carrier is insoluble, to produce an antigen-antibody reaction therebetween; (b) reacting the sample antigen with the carrier antibody for a predetermined reaction period; (c) measuring the absorbance (A) of the test dispersion after the reaction period at a first wavelength (λ
1) to determine a first absorbance value (Aλ
1), and measuring the absorbance (A) of the test dispersion after the reaction period at a second wavelength (λ
2) to determine a second absorbance value Aλ
2);(d) determining the absorbance ratio (Aλ
1 /Aλ
2) of the first absorbance value (Aλ
1) and the second absorbance value (Aλ
2) of the test dispersion; and(e) determining the concentration of the sample antigen by comparing the absorbance ratio of the test dispersion (Aλ
1 /Aλ
2) to reference absorbance ratios at said wavelengths of reference dispersions of said carrier having said antibody affixed thereto and reacted with pre-selected concentrations of said antigen. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 10)
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9. A method for determining the concentration of an antibody in a sample, comprising:
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(a) adding the sample to a test dispersion of a finely divided carrier having an antigen affixed thereto in a dispersing medium in which the carrier is insoluble, to produce an antibody-antigen reaction therebetween; (b) reacting the sample antibody with the carrier antigen for a predetermined reaction period; (c) measuring the absorbance (A) of the test dispersion after the reaction period at a first wavelength (λ
1) to determine a first absorbance value (Aλ
1), and measuring the absorbance (A) of the test dispersion after the reaction period at a second wavelength (λ
2) to determine a second absorbance value (Aλ
2);(d) determining the absorbance ratio of the first absorbance value (Aλ
1) and the second absorbance value (Aλ
2) of the test dispersion; and(e) determining the concentration of the sample antibody by comparing the absorbance ratio of the test dispersion (Aλ
1 /Aλ
2) to reference absorbance ratios at said wavelengths of reference dispersions of said carrier having said antigen affixed thereto and reacted with pre-selected concentrations of said antibody. - View Dependent Claims (11, 12, 13, 14, 15, 16)
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Specification