A kind of preparation method of leucine aminopeptidase quality control product

A technology of leucine amino group and quality control substance, applied in the field of medical testing, can solve the problems of difficulty in ensuring the stability of LAP, inconsistent test results, difficulty in popularization and application, etc., and achieves the effects of avoiding matrix effect, low production cost and easy acquisition.

Active Publication Date: 2017-11-28
杭州美康盛德医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection method of LAP is that the LAP in the sample undergoes catalytic hydrolysis of the substrate L-leucine-P-nitroaniline to generate leucine and p-nitroaniline. The absorbance of the reaction system increases at 405nm, and the rate of increase is The activity of LAP in the sample can be obtained. LAP detection kits using this method have appeared at home and abroad, but due to the lack of international standards, the detection results may be inconsistent with different manufacturers and reagents, and it is difficult to guarantee the LAP The stability of the determination has brought great difficulties to the promotion and application of the project

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Collect 150ml of serum from healthy people who have excluded infectious diseases and common diseases, and use it as a serum raw material, and place it in room temperature and -20°C refrigerators at intervals of 24 hours to repeatedly thaw and freeze, and carefully separate out after 20 days For the upper serum, first filter once with dense filter paper, then filter once with 0.45 μm filter membrane and 0.22 μm filter membrane respectively, and adjust the pH to 7.0;

[0023] (2) Take 100ml of the above-mentioned serum whose pH value has been adjusted, add 10mmol / L potassium maleate, mix well, then add 0.2g / L concanavalin A, mix well;

[0024] (3) Take 100ml of the above (2) solution, add 20mmol / L glycine, 5g / L polyethylene glycol 8000, 5g / L bovine serum albumin, 0.3g / L sodium azide, and mix well;

[0025] (4) 1ml / bottle, freeze-dried.

Embodiment 2

[0027] (1) Collect 150ml of serum from healthy people who have excluded infectious diseases and common diseases, and use it as a serum raw material, and place it in room temperature and -20°C refrigerators at intervals of 24 hours to repeatedly thaw and freeze, and carefully separate out after 30 days For the upper serum, first filter once with dense filter paper, then filter once with 0.45 μm filter membrane and 0.22 μm filter membrane respectively, and adjust the pH to 7.5;

[0028] (2) Take 100ml of the serum whose pH value has been adjusted, add 20mmol / L sodium maleate, mix well, then add 0.3g / L concanavalin A, mix well;

[0029] (3) Take 100ml of the above (2) solution, add 50mmol / L glutamic acid, 8g / L polyethylene glycol 3400, 8g / L bovine serum albumin, 0.3g / L sodium azide, and mix well;

[0030] (4) 1ml / bottle, freeze-dried.

[0031] The results of quality control products prepared in the above embodiments of the present invention stored in an environment of 2-8°C for ...

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Abstract

The invention discloses a preparation method of a leucine aminopeptidase quality control product, which comprises: using healthy human serum, freezing and thawing for 10-15 times, filtering, and the pH value of the filtrate being 7.0-7.5; adding maleate, and mixing uniformly; Then add concanavalin A, mix well; in the final mixture, the maleate concentration is 5~20mmol / L, the concentration of concanavalin A is 0.1~0.4g / L; Add amino acid in above-mentioned mixed solution, Polyethylene glycol, bovine serum albumin, preservative, mixed evenly; amino acid 10-100mmol / L, polyethylene glycol 1-10g / L, bovine serum albumin 1-10g / L, preservative in the final mixture obtained 0.2‑0.5g / L; aliquoted, freeze-dried. The invention has the advantage of ensuring the stability of LAP determination.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a preparation method of a leucine aminopeptidase quality control product. Background technique [0002] Leucine aminopeptidase (LAP, EC 3.4.11.1) is a proteolytic enzyme that is widely distributed in liver, pancreas, kidney and other tissues. When these tissues have lesions, serum LAP levels can increase. high. At present, the determination of serum LAP is an index for the diagnosis of liver diseases, and it is valuable for the differential diagnosis of cholestatic hepatitis, biliary obstruction and pancreatic cancer. Acyltransferases reflect liver disease more sensitively. The detection method of LAP is that the LAP in the sample undergoes catalytic hydrolysis of the substrate L-leucine-P-nitroaniline to generate leucine and p-nitroaniline. The absorbance of the reaction system increases at 405nm, and the rate of increase is The activity of LAP in the sample can be ob...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28G01N33/573
Inventor 沃燕波邹炳德邹继华
Owner 杭州美康盛德医学检验实验室有限公司
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