Title:
ANTI-HUMAN VITRONECTIN/THROMBIN/ANTITHROMBIN III COMPLEX MONOCLONAL ANTIBODY, HYBRIDOMA AND IMMUNOASSAY
Document Type and Number:
Japanese Patent JP3841364
Kind Code:
B2
Abstract:
PURPOSE: To obtain the subject new antibody for diagnosing blood coagulation blood coagulation sthenia, accentuation, specifically reacting with vitronectin/ thrombin/antithrombin III complex (VTAT), not reacting with each liberated material, capable of measuring VTAT in high sensitivity.
CONSTITUTION: Vitronectin is reacted with thrombin and antithrombin III in 1M tris hydrochloric acid buffer solution (pH7.5) containing 0.9% NaCl at 37°C for 30 minutes and purified by molecular sieve chromatography to obtain VTAT. Then VTAT and an adjuvant are administered to the abdominal cavity of a mouse and the mouse is immunized. After the final immunization, splenic cells are collected, fused with mouse myeloma cells and selectively cultured in a HAT medium to give a hybridoma. Then the hybridoma is cloned by a limiting dilution method to establish an antibody forming strain. The antibody forming strain is cultured to give the VTAT monoclonal antibody which specifically reacts with VTAT but not with liberated vitronectin, thrombin and antithrombin III.
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Inventors:
Xu Yoshio
Isao Kono
Kimiko Inuzuka
Yumiko Ito
Isao Kono
Kimiko Inuzuka
Yumiko Ito
Application Number:
JP27827894A
Publication Date:
November 01, 2006
Filing Date:
October 18, 1994
Export Citation:
Assignee:
Mitsubishi Chemical Yatron Co., Ltd.
International Classes:
C07K16/36; G01N33/53; C07K14/745; C07K14/78; C07K16/00; C07K19/00; C12N5/10; C12N15/02; C12P21/08; G01N33/533; G01N33/577; C12R1/91; (IPC1-7): C07K19/00; C07K14/745; C07K14/78; C12N5/10; C12N15/02; C12P21/08; G01N33/53; G01N33/533; G01N33/577; //(C12P21/08; C12R1:91)
Domestic Patent References:
| JP62138187A |
Other References:
PREISSNER, K.T.,Self-Association of Antithrombin III Relates to Multimer Formation of Thrombin- Antithrombin III Com,Thrombosis and Haemostasis,1993年,Vol.69, No.5,pp.422-429
Amiral, J. et al.,Assay of Blood Activation by Measurement of AT III-Serine Esterase Complexes using a Specific Monocl,Thrombo Haemostas,1989年,Vol.62,pp.479
TOMASINI, B.R. et al.,Coformational States of Vitronectin:Preferential Expression of an Antigenic Epitope when Vitronecti,Blood,1988年,Vol.72, No.3,pp.903-912
HAYMAN, E.G. et al.,Serum spreading factor (vitronectin) is present at the cell surface and in tissues,Proc.Natl.Acad.Sci.USA.,1983年,Vol.80,pp.4003-4007
Amiral, J. et al.,Assay of Blood Activation by Measurement of AT III-Serine Esterase Complexes using a Specific Monocl,Thrombo Haemostas,1989年,Vol.62,pp.479
TOMASINI, B.R. et al.,Coformational States of Vitronectin:Preferential Expression of an Antigenic Epitope when Vitronecti,Blood,1988年,Vol.72, No.3,pp.903-912
HAYMAN, E.G. et al.,Serum spreading factor (vitronectin) is present at the cell surface and in tissues,Proc.Natl.Acad.Sci.USA.,1983年,Vol.80,pp.4003-4007
Attorney, Agent or Firm:
Kenichi Morita