Method for measuring phosphorylated tau protein
Disclosed is a method for measuring phosphorylated tau protein in a biological sample collected from a subject, comprising the steps of: preparing a measurement sample containing a non-capture bead and a capture bead to which an immune complex is bound, by forming the immune complex of the phosphorylated tau protein in the biological sample, a capture antibody and a detection antibody on the capture bead, in the presence of the non-capture bead; and detecting a signal derived from the immune complex in the measurement sample.
1. A method for measuring phosphorylated tau protein in a biological sample collected from a subject, comprising the steps of:
preparing a measurement sample by forming an immune complex of the phosphorylated tau protein in the biological sample, a capture antibody and a detection antibody on a capture bead, in the presence of a non-capture bead, wherein the measurement sample comprises the non-capture bead to which the immune complex does not bind and the capture bead to which the immune complex is bound, and wherein the capture antibody is not immobilized on the non-capture bead;
detecting a signal derived from the immune complex in the measurement sample; and
comparing said detected signal to a reference value, and determining that said test sample is from a subject with Alzheimer's disease based on the comparison,
wherein at least one of the detection antibody and the capture antibody specifically recognizes the phosphorylated tau protein,
an epitope of the capture antibody is different from an epitope of the detection antibody, and
the number ratio of the capture beads and the non-capture beads mixed in the preparation step is 4 to 9 of the non-capture beads to 1 of the capture beads.
2. The method according to claim 1 , wherein in the detection step, the phosphorylated tau protein is measured based on the signal.
3. The method according to claim 1 , wherein, in the detection step,
the measurement sample is brought into contact with a substrate having a plurality of microwells,
the capture beads and the non-capture beads in the measurement sample are arranged one by one in the microwells, and
a signal derived from the immune complex on the capture beads arranged in the microwells is detected,
wherein the microwells have a volume capable of storing single capture bead or single non-capture bead.
4. The method according to claim 3 , wherein in the detection step,
the number of microwells comprising the capture bead on which the immune complex is formed or the number of capture beads on which the immune complex is formed in the microwell is calculated,
the phosphorylated tau protein is measured based on the calculation result.
5. The method according to claim 1 , wherein, in the detection step, signal detection is performed by imaging the signal derived from the immune complex.
6. The method according to claim 1 , wherein the biological sample is a blood sample.
7. The method according to claim 6 , wherein the blood sample is serum or plasma.
8. The method according to claim 1 , wherein the immune complex is formed by mixing a capture antibody bound to the capture bead, a biological sample containing phosphorylated tau protein, and a detection antibody, in the presence of the non-capture bead.
9. The method according to claim 1 , wherein the signal is a fluorescent signal.
10. The method of claim 1 , wherein the capture antibody is able to specifically bind to the capture bead, and wherein the capture antibody is unable to specifically bind to the non-capture bead.
11. A method for measuring phosphorylated tau protein in a biological sample collected from a subject, comprising the steps of:
preparing a measurement sample by forming an immune complex of the phosphorylated tau protein in the biological sample, a capture antibody and a detection antibody on a capture bead, in the presence of a non-capture bead, wherein the measurement sample comprises the non-capture bead to which the immune complex does not bind and the capture bead to which the immune complex is bound;
detecting a signal derived from the immune complex in the measurement sample; and
comparing said detected signal to a reference value, and determining that said test sample is from a subject with Alzheimer's disease based on the comparison,
wherein the non-capture bead is a bead on which the capture antibody is not immobilized,
at least one of the detection antibody and the capture antibody specifically recognizes the phosphorylated tau protein, and
the number ratio of the capture beads and the non-capture beads mixed in the preparation step is 4 to 9 of the non-capture beads to 1 of the capture beads.
12. The method according to claim 11 , wherein the biological sample is serum or plasma.
13. The method of claim 11 , wherein the capture antibody is able to specifically bind to the capture bead, and wherein the capture antibody is unable to specifically bind to the non-capture bead.