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Purification of human myocardium troponin I and preparation method of its monoclonal anti-body

A cardiac troponin, monoclonal antibody technology, applied in the direction of anti-animal/human immunoglobulin, animal/human peptide, peptide source, etc., can solve the problems of time for diagnosis and treatment, unfavorable disease patients, high price, etc. Achieve the effect of fast measurement time, easy promotion and application, and reduction of expensive costs

Inactive Publication Date: 2004-05-12
上海润东生物科技有限公司
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Problems solved by technology

In foreign countries, the immunoenzyme detection method is mainly completed by using automatic analyzers (such as Status, Access and other automatic analyzers), and the results can be obtained within 1 hour, but it is necessary to purchase supporting reagents such as cellulose acetate membranes coated with antibodies, and this type of Reagents are expensive
ELISA double-sandwich quantitative method is also used for detection, but this method is generally operated manually and takes a long time, which is not conducive to gaining valuable time for diagnosis and treatment of patients with diseases

Method used

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Embodiment Construction

[0026] Below in conjunction with embodiment above-mentioned method of the present invention is described in further detail:

[0027] (1) the present invention to the purification method of human cardiac troponin I:

[0028]1. Purification of human cardiac troponin C (cTnC)

[0029] (1) Take 100g of human ventricular muscle tissue fresh or stored at -75°C after liquid nitrogen quick-frozen, remove fat and fibers and shred, add 3 to 5 times the volume of homogenization buffer (Tris-HCl 50mmol / L, KCl 50mmol / L L, EDTA 1mmol / L, pH7.5) was fully homogenized, centrifuged for 30 minutes, the supernatant was removed, the precipitate was taken, and the above steps were repeated 10 times; fully homogenized each time. The precipitate was extracted more than 2 times with ethanol and ether, and dried into a dry powder. Dissolve the dry powder in buffer solution 1mol / L KCl for 12-24 hours, centrifuge at 10000g for 30 minutes, remove the precipitate, and take the supernatant;

[0030] (2) ...

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Abstract

The present invention discloses a method for purifying human myocardium troporin I, preparing its monoclonal antibody and determining its content by using one-step process. It adopts affinity chromatography to purity human myocardium troponin I, and utilizes the purified myocardium troponin I and BALB / c mouse to immunoprepare the monoclonal antibody of said myocarbium troponin I, and establishes a method for determining myocardium troponin I content by means of one-step process. It can be extensively used for diagnosing heart disease.

Description

technical field [0001] The invention relates to a method for purifying human cardiac troponin I by affinity chromatography, a method for preparing a monoclonal antibody of the cardiac troponin I and a method for determining its content by one-step method. Background technique [0002] Myocardial ischemic injury, especially acute myocardial infarction (AMI) is one of the major diseases that threaten human life. Clinically, efforts have been made to find serum indicators with good specificity and high sensitivity to diagnose it. At present, creatine kinase isoformic acid (CK-MB) has been used as an important indicator for the diagnosis of myocardial infarction, but CK-MB is not unique to the heart, and also exists in a small amount in skeletal muscle of normal people. Cardiac surgery or skeletal muscle injury patients often have increased CK-MB, which can easily lead to false positive diagnosis of AMI. Recent studies have shown that cardiac troponin I (cardiac troponin I, cT...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K16/18C12P21/08
Inventor 张寄南苏恩本
Owner 上海润东生物科技有限公司
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