Method of measuring ceruloplasmin concentration in a blood spot, kit and method of diagnosing Wilson's disease
AssigneeZenovac, Inc.
InventorsSi-Houn Hahn, Young-Ju Jang, Soo-Young Lee, Ha-Cheol Shin, Sun-Young Park, Eun-Sun Yu, Hee-Sung Han
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Abstract
The present invention relates to a method for measuring a holoceruloplasmin concentration, and more particularly, to a method for measuring a holoceruloplasmin in a blood spot based on a standard concentration curve obtained through an enzyme-linked immunosorbent assay (ELISA) or a dissociation-enhanced time-resolved fluoroimmunoassay using a holoceruloplasmin-specific polyclonal antibody and a holoceruloplasmin-specific monoclonal antibody.
Claims29
A method of measuring a holoceruloplasmin concentration in a blood spot comprising:
- a) binding the holoceruloplasmin eluted from a blood spot to a holoceruloplasmin-specific polyclonal antibody and a holoceruloplasmin-specific monoclonal antibody:
- and
- b) measuring a holoceruloplasmin concentration in the blood spot based on an absorbance standard curve obtained through an assay using a holoceruloplasmin-specific polyclonal antibody and a holoceruloplasmin-specific monoclonal antibody.
A method of measuring a holoceruloplasmin concentration in a blood spot according to an enzyme-linked immunosorbent assay, the method comprising the steps of:
- manufacturing a ceruloplasmin-specific polyclonal antibody;
- manufacturing a ceruloplasmin-specific monoclonal antibody;
- conjugating horseradish peroxidase on the ceruloplasmin-specific polyclonal antibody and the ceruloplasmin-specific monoclonal antibody;
- manufacturing a standard blood spot and a control reference blood spot by removing a ceruloplasmin from a blood sample and adding a purified ceruloplasmin solution containing a holoceruloplasmin at a constant concentration to the blood sample;
- drawing out an absorbance standard curve based on the standard blood spot and the control reference blood spot using the ceruloplasmin-specific polyclonal antibody and the ceruloplasmin-specific monoclonal antibody;
- and
- measuring a holoceruloplasmin concentration in a blood spot of a patient using the standard curve through an enzyme-linked immunosorbent assay.
A method of measuring a holoceruloplasmin concentration in a blood spot according to a dissociation-enhanced time-resolved fluoroimmunoassay, the method comprising the steps of:
- manufacturing a ceruloplasmin-specific polyclonal antibody;
- manufacturing a ceruloplasmin-specific monoclonal antibody;
- conjugating with europium on the ceruloplasmin-specific polyclonal antibody and the ceruloplasmin-specific monoclonal antibody;
- manufacturing a standard blood spot and a control reference blood spot by removing a ceruloplasmin from a blood sample and adding a purified ceruloplasmin solution containing a holoceruloplasmin at a constant concentration to the blood sample;
- drawing out an fluorescence standard curve based on the standard blood spot and the control reference blood spot using the ceruloplasmin-specific polyclonal antibody and the ceruloplasmin-specific monoclonal antibody;
- and
- measuring a holoceruloplasmin concentration in a blood spot of a patient using the standard curve through a dissociation-enhanced time-resolved fluoroimmunoassay.
A Wilson's disease screening kit reagent, comprising a holoceruloplasmin-specific polyclonal antibody, a holoceruloplasmin-specific monoclonal antibody, a standard blood spot, and a control reference blood spot.
A Wilson's disease screening kit, for measuring a holoceruloplasmin concentration in a blood spot based on an absorbance standard curve obtained through an enzyme-linked immunosorbent assay using a holoceruloplasmin-specific polyclonal antibody, a holoceruloplasmin-specific monoclonal antibody, a standard blood spot, and a control reference blood spot.
A Wilson's disease screening kit, for measuring a holoceruloplasmin concentration in a blood spot based on a fluorescence standard curve obtained through a dissociation-enhanced time-resolved fluoroimmunoassay using a holoceruloplasmin-specific polyclonal antibody, a holoceruloplasmin-specific monoclonal antibody, a standard blood spot, and a control reference blood spot.
Dependencies are inferred from the claim text.