| CPC G01N 21/6445 (2013.01) [G01N 33/582 (2013.01); G01N 21/64 (2013.01); G01N 21/6428 (2013.01); G01N 33/53 (2013.01)] | 6 Claims |
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1. A fluorescence polarization immunoassay method for using an antibody that has binding ability to a target substance and a fluorescently labeled substance, obtained by labeling the target substance with a fluorochrome, to measure a concentration of the target substance included in a first sample to be measured, the method comprising:
measuring a degree of polarization of a first reference sample that does not include the target substance;
measuring a degree of polarization of each of a plurality of second reference samples that are produced by adding the antibody, the fluorescently labeled substance, and the target substance to the first reference sample, and that have mutually different concentrations of the target substance;
measuring a degree of polarization of the first sample to be measured;
measuring a degree of polarization of a second sample to be measured that is produced by adding the antibody and the fluorescently labeled substance to the first sample to be measured in amounts equal to those added to the second reference samples;
calculating a first corrected degree of polarization by correcting the degree of polarization of the second reference samples with the degree of polarization of the first reference sample, and generating a first calibration curve expressing a relationship between the concentration of the target substance included in the second reference samples and the first corrected degree of polarization;
calculating a second corrected degree of polarization by correcting the degree of polarization of the second sample to be measured with the degree of polarization of the first sample to be measured; and
obtaining, as the concentration of the target substance included in the first sample to be measured, a concentration corresponding to the second corrected degree of polarization in the first calibration curve.
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