Cholesterol measuring reagent containing a cholesterol esterase
There are provided a method and reagent for measuring cholesterol in remnant-like lipoprotein in a sample with high sensitivity by more simple operation. The method for measuring cholesterol in remnant-like lipoprotein uses a cholesterol esterase, in which the activity ratio of a lipoprotein lipase to a cholesterol esterase (lipoprotein lipase activity/cholesterol esterase activity) is from 12 to 7000 in a method for measuring cholesterol in the lipoprotein by measuring hydrogen peroxide or a reduced coenzyme obtained by allowing the cholesterol esterase and a cholesterol oxidase or a cholesterol dehydrogenase to act on a test sample containing a lipoprotein.
1. A method for measuring cholesterol in remnant-like lipoprotein, comprising measuring cholesterol in the lipoprotein by measuring hydrogen peroxide or a reduced coenzyme obtained by allowing a cholesterol esterase and a cholesterol oxidase or a cholesterol dehydrogenase to act on a test sample containing a lipoprotein, wherein said cholesterol esterase has lipoprotein lipase activity and cholesterol esterase activity wherein the activity ratio of lipoprotein lipase activity to cholesterol esterase activity ranges from 12:1 to 7000:1.
2. The method according to claim 1 , wherein the activity ratio of lipoprotein lipase activity to cholesterol esterase activity ranges from 28:1 to 800:1.
3. The method according to claim 1 , wherein the cholesterol esterase is selected from a lipase from Chromobacterium viscosum wherein said activity ratio ranges 460:1 to 800:1 identified as LP, a lipase from an unknown microorganism wherein said activity ratio ranges from 5600:1 to 6900:1 identified as LPBP, a cholesterol esterase from Pseudomonas sp. wherein said activity ratio ranges from 13:1 to 16:1 identified as CEN, a cholesterol esterase from Pseudomonas sp. wherein said activity ratio ranges from 13:1 to 16:1 identified as CE, and a cholesterol esterase from Pseudomonas sp. wherein said activity ratio ranges from 28:1 to 35:1 identified as COE 311.
4. The method according to any one of claims 1 to 3 , further comprising pretreating the test sample with an anti-apoB-100 antibody and/or an anti-apoA-I antibody.
5. A measuring reagent for cholesterol in remnant-like lipoprotein, comprising:
a cholesterol esterase having lipoprotein lipase activity and cholesterol esterase activity wherein the activity ratio of lipoprotein lipase activity to cholesterol esterase activity ranges from 12:1 to 7000:1; and
a cholesterol oxidase or a cholesterol dehydrogenase.
6. The measuring reagent according to claim 5 , wherein the activity ratio of lipoprotein lipase activity to cholesterol esterase activity ranges from 28:1 to 800:1.
7. The reagent according to claim 5 , wherein the cholesterol esterase is selected from a lipase from Chromobacterium viscosum wherein said activity ratio ranges 460:1 to 800:1 identified as LP, a lipase from an unknown microorganism wherein said activity ratio ranges from 5600:1 to 6900:1 identified as LPBP, a cholesterol esterase from Pseudomonas sp. wherein said activity ratio ranges from 13:1 to 16:1 identified as CEN, a cholesterol esterase from Pseudomonas sp. wherein said activity ratio ranges from 13:1 to 16:1 identified as CE, and a cholesterol esterase from Pseudomonas sp. wherein said activity ratio ranges from 28:1 to 35:1 identified as COE 311.
8. A measuring reagent for cholesterol in remnant-like lipoprotein, comprising;
a first reagent comprising an anti-apoB-100 antibody and/or an anti-apoA-I antibody; and
a second reagent comprising the measuring reagent according to any one of claims 5 to 7 .