Method of selectively assaying adiponectin multimers
View Patent ↗Kits and methods for selectively assaying a target adiponectin multimer in a biological sample. Such methods accurately evaluate the relationship between a disease and adiponectin through selective assay of adiponectin multimers and provide information that cannot be obtained through measurement of the total amount of adiponectin alone. A method for selectively assaying a target adiponectin multimer in a biological sample comprising distinguishing target adiponectin multimer from the other adiponectin multimers by using a protease and/or an antibody.
1. A kit comprising:
(a) a protease which digests ULMW-Ad (ultra low molecular weight adiponectin), LMW-Ad (low molecular weight adiponectin) and MMW-Ad (middle molecular weight adiponectin) multimers; and
(b) an anti-human adiponectin antibody which recognizes adiponectin and which is immobilized on an insoluble carrier.
2. The kit of claim 1 , wherein the protease is derived from a microorganism.
3. The kit of claim 1 , wherein the protease is selected from at least one of proteinase K, protease P “Amano,” protease N “Amano”, and Umamizyme.
4. The kit of claim 1 , wherein the at least one protease includes proteinase K.
5. The kit of claim 1 , wherein the anti-human adiponectin antibody is polyclonal.
6. The kit of claim 1 , wherein the anti-human adiponectin antibody is monoclonal.
7. The kit of claim 1 , wherein the anti-human adiponectin antibody is polyclonal.
8. The kit of claim 1 , wherein the anti-human adiponectin antibody is a goat, mouse or rabbit antibody.
9. The kit of claim 1 , further comprising an enzyme-labeled antibody that recognizes albumin in combination with the carrier.
10. The kit of claim 1 , wherein the anti-human adiponectin antibody recognizes an adiponectin-albumin complex.
11. The kit of claim 1 , which is an ELISA (enzyme linked immunosorbent assay) kit.
12. The kit of claim 1 , which is a kit for CLEIA (chemiluminescent enzyme immunoassay), RIA (radioimmunoassay), or LTIA (latex turbidimetric immunoassay).
13. The kit of claim 1 , further comprising at least one of a reducing agent, an acid or a salt thereof, a surfactant, a stabilizer, or a buffer.
14. The kit of claim 1 , further comprising a biological sample suspected of containing adiponectin.
15. A composition comprising:
(a) a protease which digests ULMW-Ad (ultra low molecular weight adiponectin), LMW-Ad (low molecular weight adiponectin) and MMW-Ad (middle molecular weight adiponectin); and
(b) an anti-human adiponectin antibody which recognizes adiponectin and which is immobilized on an insoluble carrier.
16. The composition of claim 15 , further comprising at least one of a reducing agent, an acid or a salt thereof, a surfactant, or a buffer.
17. The composition of claim 15 , further comprising a biological sample suspected of containing adiponectin.