Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1998-01-14
1999-10-26
Eisenschenk, Frank C.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 5, 435 71, 435 79, 530300, 530328, 530344, G01N 3353, G01N 33542, C12Q 170, A61K 3800
Patent
active
059726334
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to antibodies against heart muscle actin, processes for the production thereof and their use.
Processes are known for detecting a cardiac infarct by determining the heart muscle-specific creatine phosphokinase and by determining the serum-lactate dehydrogenase, respectively. However, the former determination cannot be made until 3 hours following a cardiac infarct. Also, only a maximum of 50% of the creatine phosphokinase is present after one day. Furthermore, the serum-lactate dehydrogenase is not a heart-specific enzyme. Thus, the above determinations are only suitable in a limited fashion for detecting a cardiac infarct or another lesion of the heart muscle.
Therefore, it is the object of the present invention to provide a product by which a cardiac infarct or another lesion of the heart muscle can be detected specifically and reliably.
According to the invention this is achieved by antibodies which are directed against heart muscle actin.
It follows from applicant's experiments that in the case of cardiac infarct heart muscle actin is released. This actin differs as regards few amino acids from the actin of the sceletal muscle and that of the smooth muscles, respectively (cf. Vandekerckhove, J. and Weber, K., Differentiation 14 (1979), pp. 123-133). According to the invention these differences are used to direct antibodies against heart muscle actin.
Such antibodies may be polyclonal or monoclonal, monoclonal antibodies being preferred. The antibodies may have been obtained from any animal or human being, rabbits being preferred for polyclonal antibodies and mice being preferred for monoclonal antibodies.
In addition, the antibodies may be synthetic, portions which are not necessary for detecting heart muscle actin being optionally lacking fully or partially therefrom and these portions being replaced by others which provide the antibodies with further favorable properties, respectively.
The expression "heart muscle actin" comprises heart muscle actin of any kind and origin as well as fragments thereof. It may originate from human beings or animals. In addition, the heart muscle actin may be present in free or complexed form.
Preferred antibodies of the present invention, namely the monoclonal mouse antibodies Ac1-12.3.1 and Ac1-20.4.2 were deposited with DSMZ located at Mascheroder Weg 1b 38124 Braunschweig, Germany [German-Type Collection of Microorganisms] under numbers DSM ACC 2208 and DSM ACC 2209, respectively, on Mar. 22, 1995 in accordance with the terms of the Budapest Treaty.
Antibodies according to the invention may be produced according to conventional processes. If polyclonal antibodies and monoclonal antibodies, respectively, shall be produced, it will be favorable to immunize animals, particularly rabbits for the former antibodies and mice for the latter antibodies, with an above heart muscle actin and/or fragments thereof, particularly fragments of the N-terminus or C-terminus of heart muscle actin. The expression "fragments thereof" also comprises synthetic peptides which include partial sequences, particularly of the N-terminus or C-terminus, of heart muscle actin. It may also be advantageous to immunize the animals with a mixture consisting of heart muscle actin and/or fragments thereof. Further boosters of the animals may take place with the same heart muscle actin or actins and/or fragments thereof. It is also possible to use other heart muscle actins and/or fragments thereof or a combination of these and the preceding heart muscle actin or actins and/or fragments thereof for booster.
The polyclonal antibodies may then be obtained from the serum of the animals. Spleen cells from the animals are fused with myeloma cells for the monoclonal antibodies.
For the production of synthetic antibodies, e.g. the above obtained monoclonal antibodies may be used as a basis. For this purpose, it is an obvious thing to analyze the antigen binding regions of the monoclonal antibodies and identify the portions necessary and not necessary for the specific heart muscle a
REFERENCES:
Aranega, et al: Circulating alpha-actin protein in acute myocardial infaction: Int. J. Car.: Bd. 38, Nr. 1: pp. 49-55, Jan. 1993.
Hanin, et al. : Production of oligoclonal antibodies . . . : Pep. Res. : vol. 2, No. 6 :pp. 367-372, 1989.
International Journal of Cardiology, Bd. 38, Nr. 1, Jan. 1993, Amsterdam, NL, Seiten 49-55, XP00578319, A.E. Aranega et al, "Circulating Alpha-Actin Protein In Acute Myocardial Infarction".
Journal of Molecular and Cellular Cardiology, Bd. 25, Nr. 1, Jan. 1993, London, GB, Seiten 15-22, XP000578297, A.E. Aranega et al, "Circulating Alpha-Actin In Angina Pectoris".
Peptide Research, Bd. 2, Nr. 6, Nov. 1989, Natrick, GB, Seiten 367-372, XP002010880, V. Hanin et al, "Production of Oligoclonal Antibodies Directed to the N-Terminal of Smooth Muscle Alpha Actin Using Peptidyl-Polyacrylic Resins as Direct Immunogens".
"Sigma Immuno Chemicals, 1992 Katalog", 1992, Sigma Chemie GmbH, Deisenhofen, DE, XP002010881.
Franke Werner Wilhelm
Stehr Sabine
Stumpp Sabine
Eisenschenk Frank C.
Nelson Brett
Progen Biotechnik GmbH
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