Claims
- 1. A method of immunologically assaying the total amount of human osteocalcin and its fragment in a human assay sample using a solid-phase antibody and a labelled antibody, which comprises:
- bringing the human assay sample into contact with a solid-phase antibody and a labelled antibody,
- reacting the sample with the solid-phase and labelled antibodies so as to form a complex of the labelled antibody:human osteocalcin or its fragment:solid-phase antibody in the human sample, and
- detecting the amount of labelled antibody in the complex, wherein
- (1) one of the antibodies is a monoclonal antibody or antibody fragment to human osteocalcin which specifically binds to a first epitope within a region of amino acids 1 to 20 of the N-terminal of human osteocalcin, and
- (2) the other antibody is a polyclonal antibody or antibody fragment to human osteocalcin which specifically binds to a second epitope within said region of amino acids 1 to 20 of the N-terminal of human osteocalcin.
- 2. The method of claim 1 wherein the solid-phase antibody is the polyclonal antibody or antibody fragment and the labelled antibody is the monoclonal antibody or antibody fragment.
- 3. The method of claim 1 wherein the labelled antibody is an Fab' or F(ab').sub.2 antibody fragment.
- 4. The method of claim 1 wherein the labelled antibody is labelled with an enzyme.
- 5. The method of claim 1 wherein the solid-phase antibody is bound to a plastic bead having a smooth surface with a centerline average roughness (Ra) of not more than 1.5 micrometers.
- 6. The method of claim 1 wherein the solid-phase antibody is the monoclonal antibody or antibody fragment and the labelled antibody is the polyclonal antibody or antibody fragment.
- 7. A reagent for immunologically assaying the total amount of intact human osteocalcin and its fragment in a human assay sample, comprising a solid-phase antibody and a labelled antibody, wherein
- (1) one of the antibodies is a monoclonal antibody or antibody fragment to human osteocalcin which specifically binds to a first epitope within a region of amino acids 1 to 20 of the N-terminal of human osteocalcin, and
- (2) the other antibody is a polyclonal antibody or antibody fragment to human osteocalcin which specifically binds to a second epitope within said region of amino acids 1 to 20 of the N-terminal of human osteocalcin.
- 8. A kit for immunologically assaying the total amount of intact human osteocalcin and its fragment wherein the kit is composed of:
- (a) a solid-phase antibody,
- (b) a labelled antibody,
- (c) a dilution buffer, and
- (d) a washing agent,
- wherein
- (1) one of the antibodies is a monoclonal antibody or antibody fragment to human osteocalcin which specifically binds to a first epitope within a region of amino acids 1 to 20 of the N-terminal of human osteocalcin, and
- (2) the other antibody is a polyclonal antibody or antibody fragment to human osteocalcin which specifically binds to a second epitope within said region of amino acids 1 to 20 of the N-terminal of human osteocalcin.
- 9. The kit according to claim 8, wherein the label is an enzyme, and wherein the kit further includes a substrate for measuring the enzyme activity and a reaction stopping agent.
Priority Claims (2)
Number |
Date |
Country |
Kind |
1-30003 |
Feb 1989 |
JPX |
|
1-255306 |
Oct 1989 |
JPX |
|
Parent Case Info
This is a divisional application of Ser. No. 08/316,364, filed Oct. 3, 1994, now U.S. Pat. No. 5,506,111, which is a continuation application of Ser. No. 08/038,544 filed Mar. 29, 1993, now abandoned, which is a continuation application of Ser. No. 07/582,849 filed as PCT/JP90/00155 Feb. 8, 1990, now abandoned.
US Referenced Citations (2)
Number |
Name |
Date |
Kind |
4438208 |
Deftos et al. |
Mar 1984 |
|
5506111 |
Hosoda et al. |
Apr 1996 |
|
Divisions (1)
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Number |
Date |
Country |
Parent |
316364 |
Oct 1994 |
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Continuations (2)
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Number |
Date |
Country |
Parent |
38544 |
Mar 1993 |
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Parent |
582849 |
Oct 1990 |
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