Section of synthesized peptide S28-42 of troponin I of human cardiac muscle and application

A troponin, synthetic peptide technology, applied in the direction of peptides, material testing products, instruments, etc., to achieve the effect of avoiding weak immunogenicity, high detection level, strong immunogenicity and immunoreactivity

Active Publication Date: 2005-08-31
江苏如东高新创业投资有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the amino terminal and carboxyl terminal of human cTnl are the strongest epitope regions, this region is also sensitive to oxidation, reduction and phosphorylation, and is easily hydrolyzed by endogenous proteolytic enzymes. Different states have different immune responses. active

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0021] 1. Using synthetic peptide S 28-42 MBS (m-maleimide benzoic acid-N-hydroxysuccinimide ester) method coupled with carrier protein, due to S 28-42 Due to the difference in sequence with human skeletal muscle troponin and mouse cardiac troponin, a higher immune effect can be achieved after immunizing BALB / c mice. The specific method is as follows:

[0022] 1) m-maleimide benzoic acid-N-hydroxysuccinimide ester (MBS) was prepared into 5 mg / ml dimethylformamide (DMF), and prepared within one hour before use.

[0023] 2) Dissolve 10 mg of keyhole limpet hemocyanin (KLH) in 1 ml of deionized water.

[0024] 3) Put 1ml of KLH solution into a small test tube, and add 140ul of DMF solution dissolved in 2mg of MBS. Stir at room temperature for 30 min. Pass through a G-25 column (PBS, pH 7.4) to collect the gray product. 5 mg of polypeptide was dissolved in not more than 0.5 ml of PBS (0.01 mol / L) at pH 7.4. The two were mixed and stirred at room temperature for 2 hours. Dia...

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PUM

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Abstract

A synthetic peptide fragment S27-42 of human myocardium troponin I (cTnI) for testing the damage of human myocardium is disclosed. It has strong immunogenicity and immunoreactivity. Using it to immunize vertebrates can generate high-specificity antibody of cTnI to detect the similar peptide fragment residue in serum after cTnI is degradated. It can be used to prepare the reagent kit to detect the myocardium damage.

Description

technical field [0001] The invention relates to a polypeptide which can be used for producing and screening high-efficiency antiserum and highly specific monoclonal antibody against a specific antigenic determinant epitope of human cardiac troponin I, and the application of the polypeptide. Background technique [0002] Myocardial ischemic injury, especially acute myocardial infarction (AMI) is one of the major diseases that threaten human life. People have been working hard to find indicators with good specificity and high sensitivity for the diagnosis of AMI. With the in-depth study of myocardial markers, biochemical markers have been developed from the earlier aspartate aminotransferase (AST), serum lactate dehydrogenase (LD) and isoenzymes, creatine kinase (CK) and isoenzymes So far, the diagnostic sensitivity and specificity of various early markers (such as myoglobin, etc.) and specific markers (cardiac troponin) have been enhanced. [0003] Cardiac troponin I (cTnI) ...

Claims

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Application Information

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IPC IPC(8): C07K7/08G01N33/577G01N33/68
Inventor 朱乃硕黄应峰
Owner 江苏如东高新创业投资有限公司
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