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Method for inhibiting protease in biological sample containing pancreatic juice component

A protease inhibitor, biological sample technology, applied in the direction of protease inhibitors, biochemical equipment and methods, and microbial determination/inspection, etc., can solve problems such as molecular damage, and achieve the effect of inhibiting decomposition and stable preservation

Inactive Publication Date: 2014-05-07
OLYMPUS CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cryopreserved pancreatic juice needs to be lysed before measurement, and the lysed cells, proteins, etc. may be decomposed again
Also, the molecule itself that is the object of analysis may be damaged during cryopreservation

Method used

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  • Method for inhibiting protease in biological sample containing pancreatic juice component
  • Method for inhibiting protease in biological sample containing pancreatic juice component
  • Method for inhibiting protease in biological sample containing pancreatic juice component

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] In order to investigate the protease inhibitor having the best effect on pancreatic juice, a protease inhibitor mixture having the best inhibitory effect on pancreatic juice was searched for among eight commercially available protease inhibitor mixtures. The protease inhibitor mixture used is: complete (product of Roche) (inhibitor mixture 1), Halt Protease Inhibitor Cocktail (product of Thermo) (inhibitor mixture 2), Protease In inhibitor cocktail (product of SIGMA) (inhibitor Mixture 3), Protease Inhibitor Mix (GE company product) (inhibitor mixture 4), Protease Inhibitor Cocktail set I (M ERCK company product) (inhibitor mixture 5), Protease Inhibitor Cocktail set II (ME RCK company product) (inhibitor Agent mixture 6), Protease Inhibitor Cocktail set III (product of ME RCK company) (inhibitor mixture 7), and Protease Inhibitor Cocktail (product of BioVision company) (inhibitor mixture 8).

[0083] Use "pancreatin" as a digestive enzyme prepared from porcine pancreas...

Embodiment 2

[0086] A study was conducted on how inhibition of protease activity affects the storage stability of proteins in pancreatic juice. It was investigated to what extent S100P could be preserved with and without the addition of a protease inhibitor in a solution in which S100P (calcinbindin) normally present in pancreatic juice was mixed with trypsin (simulated pancreatic juice).

[0087] Specifically, first, a 25 ng / mL S100P (standard product) solution (sample solution 1), a solution containing 25 ng / mL S100P (standard product) and 1 mg / mL trypsin (sample solution 2), and a solution containing A solution (sample solution 3) containing 25ng / mL of S100P (standard product) and 1mg / mL of trypsin and Protease Inhibitor Mix (product of GE) at twice the recommended concentration (2×), containing 25ng / mL S100P (standard product) and 1 mg / mL trypsin and complete (Roche company product) solution (sample solution 4) containing 5 times the recommended concentration (5×). Various sample solu...

Embodiment 3

[0090] Among the various protease inhibitors contained in the inhibitor mixture having the highest protease inhibitory activity in Example 1, it was examined which protease inhibitor had the highest inhibitory activity on proteases in pancreatic juice.

[0091]The serine protease inhibitors contained in Protease Inhibitor Mix (GE company product) include 4 kinds of serine protease inhibitors, aprotinin, leupeptin, PMSF, and AEBSF (concentrations are not disclosed). Therefore, it was prepared in such a way that the final concentration was the recommended maximum concentration of each protease inhibitor single agent, and for a total of 15 combinations of 4 kinds of inhibitors, pancreatin and 2 kinds of pancreatic juice samples were used for the same as in Example 1. Protease activity was measured. The protease inhibitors added to each sample solution are shown in Table 1. In addition, the recommended maximum concentration is as follows: 0.3 μM for aprotinin (product of Roche co...

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Abstract

A method for inhibiting a protease in a biological sample containing a pancreatic juice component can inhibit the enzymatic activity of the protease in the biological sample by adding a protease inhibitor having at least one sulfonyl fluoride group to the biological sample containing a pancreatic juice component. The protease inhibitor for the biological sample containing a pancreatic juice component is a compound having a sulfonyl fluoride group, has a protease inhibitory activity, and is added to the biological sample containing a pancreatic juice component for the purpose of inhibiting the protease in the biological sample. A kit for storing a biological sample containing a pancreatic juice component involves a protease inhibitor having at least one sulfonyl fluoride group, and is used for storing the biological sample containing a pancreatic juice component.

Description

technical field [0001] The present invention relates to a method for inhibiting the enzymatic activity of protease in a biological sample containing pancreatic juice components, a protease inhibitor effective for inhibiting the decomposition of components derived from pancreatic juice in pancreatic juice or duodenal juice, and a protease inhibitor containing the protease inhibitor Kit for preservation of biological samples such as pancreatic juice. [0002] This application claims priority based on Japanese Patent Application No. 2011-199357 for which it applied in Japan on September 13, 2011, and uses the content here. Background technique [0003] Body fluid is an important biological sample for knowing the state of an organ. Pancreatic juice is also an important biological sample for understanding the state of the pancreas, and can be used for cell diagnosis, bicarbonate measurement, bacterial inspection, inspection of markers composed of protein and / or nucleic acid, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/99A01N1/02C07C309/81C07C309/88
CPCA01N1/00C12N9/99C07K14/81C12Q1/37
Inventor 片冈理惠中田真理守屋奈绪
Owner OLYMPUS CORP
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