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Aspartate aminotransferase mitochondrion isozyme detection kit and application

A mitochondrial isoenzyme, aspartate amino technology, applied in the field of immunodiagnosis, can solve the problems of poor antibody specificity, high m-AST measurement results, c-AST can not be completely inhibited, etc., to achieve the effect of improving efficiency

Active Publication Date: 2019-07-16
WUHAN LIFE ORIGIN BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is also possible that the specificity of the anti-c-AST antibody is poor, and the active center of c-AST is not completely covered by the combination of the antibody and c-AST, resulting in that c-AST cannot be completely inhibited, and the m-AST measurement result is high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Embodiment 1. The detection kit prepared by the present invention

[0012] Reagent 1 includes the following components: Tris buffer 50mmol / L; L-aspartic acid 15mmol / L; lactate dehydrogenase 0.5mmol / L; goat anti-human c-AST antibody 1 > 1KU / L; goat anti-human c - AST antibody 2>1KU / L; malate dehydrogenase>0.8KU / L; reagent 2 includes the following components: α-ketoglutarate 130mmol / L; NADH 130mmol / L. The binding epitope of goat anti-human c-AST antibody 1 is NHEYLPILGLAEFRSCASRLAL; the binding epitope of goat anti-human c-AST antibody 2 is KQIASVMKHRFLFPFFDSAYQG;

Embodiment 2

[0013] Example 2. Kits prepared using polyclonal antibodies

[0014] Reagent 1 includes the following components: Tris buffer 50mmol / L; L-aspartic acid 15mmol / L; lactate dehydrogenase 0.5mmol / L; polyclonal goat anti-human m-AST antibody>1KU / L; Hydrogenase>0.8KU / L; Reagent 2 includes the following components: α-ketoglutarate 130mmol / L; NADH 130mmol / L.

Embodiment 3

[0015] Example 3. Use of monoclonal antibodies prepared at other sites as shielding agents.

[0016] Reagent 1 includes the following components: Tris buffer 50mmol / L; L-aspartic acid 15mmol / L; lactate dehydrogenase 0.5mmol / L; goat anti-human c-AST antibody 1 > 1KU / L; goat anti-human c - AST antibody 2>1KU / L; malate dehydrogenase>0.8KU / L; reagent 2 includes the following components: α-ketoglutarate 130mmol / L; NADH 130mmol / L. The binding epitope of goat anti-human c-AST antibody 1 is LRARLEA; the binding epitope of goat anti-human c-AST antibody 2 is WAIRYFVSE;

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PUM

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Abstract

The invention relates to an aspartate aminotransferase mitochondrion isozyme detection kit. The aspartate aminotransferase mitochondrion isozyme detection kit comprises the following components: the reagent 1 comprising 50 mmol / L of Tris buffer solution, 15 mmol / L of L-aspartic acid, 0.5 mmol / L of lactate dehydrogenase, more than 1 KU / L of the goat anti-human ASTs antibody, and more than 0.8 KU / Lof the malic dehydrogenase; the reagent 2 comprising 130 mmol / L of alpha-ketoglutaric acid and 130 mmol / L of NADH.

Description

technical field [0001] The invention relates to the field of immunodiagnosis, in particular to a detection kit for mitochondrial isozyme of aspartate aminotransferase and a use method thereof. Background technique [0002] Aspartate aminotransferase (AST) is widely present in various tissue cells of the human body, especially in liver cells and cardiomyocytes, so AST is often used in the detection of liver diseases and myocardial diseases. Electrophoretic analysis further divides AST into cytosolic AST (cytosolic AST, c-AST) and mitochondrial AST (mitochondrial AST, m-AST). The activity of AST in serum is the sum of the activities of c-AST and m-AST. AST, m-AST and c-AST are closely related. Currently, m-AST detection methods mainly include immunosuppressive method and enzymatic hydrolysis method, but the results of the two methods are different. [0003] This indicated that the specificity of m-AST, an immunosuppressive method, decreased when AST and m-AST increased, and...

Claims

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Application Information

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IPC IPC(8): G01N33/573
CPCG01N33/573
Inventor 华权高沈鹤霄徐春雷
Owner WUHAN LIFE ORIGIN BIOTECH LTD
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