Anti-annexin-v monoclonal antibody, process for producing...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/20 (2006.01) C07K 16/18 (2006.01) C12P 21/08 (2006.01) G01N 33/53 (2006.01) G01N 33/577 (2006.01)

Patent

CA 2180830

Using human annexin-V or human annexin-V plus dog annexin-V as antigen(s), hybridoma cell lines are prepared which are capable of producing anti-annexin-V monoclonal antibodies having a binding specificity to antigenic determinant site on annexin-V as antigenic protein and belonging to immunoglobulin G class. By the hybridoma cell lines are produced the anti-annexin-V monoclonal antibodies, with which a diagnostic agent is provided for diagnosis of myocardial infarction and angina pectoris. There is also provided diagnosis of myocardial infarction and angina pectoris using a first and a second monoclonal antibodies produced by the hybridoma cell lines to quantitate human annexin- V in a sample, in which an antigen-antibody reaction on annexin- V in the sample is caused with the first anti-annexin-V monoclonal antibody to form an annexin-V antigen/anti-annexin-V monoclonal antibody complex, the antigenic site of annexin-V of the formed annexin-V antigen/annexin-V monoclonal antibody complex is allowed to be bound with a labeled anti-annexin-V polyclonal or second monoclonal antibody so as to form a labeled form of said annexin-V antigen/anti-annexin-V monoclonal antibody complex bound with the polyclonal or the second monoclonal antibody, and the labeled form of the complex is quantitatively analyzed. 121

Cette invention se rapporte à un anticorps monoclonal antiannexine-V, qui est produit à partir d'une lignée de cellules hybridomes préparées à l'aide d'annexine V humaine seule ou combinée à une annexine V canine, qui possède une spécificité de liaison pour le site déterminant antigénique de l'annexine V, une protéine antigénique, et qui produit un anticorps monoclonal antiannexine-V appartenant à l'immunoglobuline G. L'anticorps monoclonal antiannexine-V ainsi produit est utilisé pour diagnostiquer l'infarctus du myocarde et l'angine de poitrine, et les premier et second anticorps monoclonaux antiannexine-V produits à partir des lignées de cellules hybridomes correspondantes sont utilisés pour diagnostiquer l'infarctus du myocarde et/ou l'angine de poitrine par quantification de l'annexine humaine V contenue dans des spécimens. Ce procédé de quantification consiste à effectuer une réaction antigène-anticorps entre l'annexine V humaine et le premier anticorps monoclonal antiannexine-V, afin de former un complexe entre un antigène d'annexine V humaine et un anticorps monoclonal antiannexine-V, à lier un anticorps polyclonal antiannexine-V marqué ou le second anticorps monoclonal à la fraction antigène d'annexine V du complexe antigène-anticorps formé, afin d'obtenir un conjugué marqué entre le complexe antigène-anticorps et l'anticorps polyclonal ou le second anticorps monoclonal, puis à quantifier le conjugué marqué ainsi obtenu.

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