Agkistrodon halys venom identification method applying mass spectrometry and application of method

An identification method and a mass spectrometry technique, which are applied in the field of biopharmaceuticals, can solve the problems of not yet discovered pit viper venom method, complicated and cumbersome operation process, and low detection sensitivity, so as to overcome the complex and cumbersome operation process, simple operation process and low detection sensitivity. high effect

Pending Publication Date: 2020-07-28
SHANGHAI SERUM BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the agglutination test method has the problems of low detection sensitivity, low correct detection rate and obvious cross-reaction
Some scholars have revealed that there is a serious cross-reaction in the identification of snake venom protein snake venom by ELISA, but it is very unfavorable for the identification of snake venom
Aptamer technology has the disadvantage that the operation process is very complicated and tedious, such as "A method for detecting and identifying snake venom species using aptamer technology" (Chinese invention patent application CN102323400A)
Polymerase chain reaction is to identify from the perspective of DNA, but there are few reports on related research
[0009] At present, there is no report on the identification of Agkistrodon halys venom using mass spectrometry

Method used

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  • Agkistrodon halys venom identification method applying mass spectrometry and application of method
  • Agkistrodon halys venom identification method applying mass spectrometry and application of method
  • Agkistrodon halys venom identification method applying mass spectrometry and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 A method for identifying Agkistrodon halys venom

[0047] Proceed as follows:

[0048] (1) Choose the Agkistrodon toxin purchased from Moganshan Snake Industry Co., Ltd., Deqing County, and dissolve it in solution A to prepare 60 mg / mL;

[0049] (2) Carry out SDS-PAGE electrophoresis, the protein gel concentration is 12.5%, and the formula is as follows in Table 2:

[0050] Table 2 Protein glue concentration formula

[0051]

[0052] (3) Coomassie brilliant blue staining; see SDS-PAGE electrophoresis figure 1 , 8 protein bands can be distinguished (see figure 1 Middle 1-8);

[0053] (4) Cut out the corresponding protein band, add a small amount of solution H to cover the protein band;

[0054] (5) LC-MS / MS mass spectrometry identification: the sample is separated from the mobile phase in the liquid mass spectrometer, and after being ionized, the mass analyzer of the mass spectrometer separates the ion fragments according to the mass number, and the mas...

Embodiment 2

[0067] Example 2 Application of an identification method of Agkistrodon venom in the preparation of anti-Akistrodon venom serum for the treatment of Agkistrodon venom poisoning

[0068] Proceed as follows:

[0069] (1) After dissolving Agkistrodon halys venom (purchased from Moganshan Snake Industry Co., Ltd., Deqing County) with solution A, perform SDS-PAGE electrophoresis;

[0070] (2) SDS-PAGE electrophoresis results ( figure 2 ) and existing electrophoretic patterns ( figure 1 ) comparison, confirm that the protein is 8 main bands, that is, the corresponding Agkistrodon venom protein;

[0071] (3) Immunize the horses after detoxifying formaldehyde from Agkistrodon halys venom;

[0072] (4) After the horses immune to Agkistrodon venom are collected by the plasmapheresis machine, the production of anti-Viper venom serum is carried out to obtain the stock solution and products of Anti-Viper venom serum;

[0073] The above production process of the anti-viper venom serum ...

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Abstract

The invention discloses a method for identifying agkistrodon halys venom by applying mass spectrometry. The method comprises the following steps of: (1) dissolving agkistrodon halys venom to prepare aprotein solution; (2) carrying out SDS-PAGE analysis; and (3) carrying out LC-MS/MS mass spectrometry to carry out agkistrodon halys venom component identification. In addition, the invention furtherdiscloses application of the identification method in the preparation of products for identifying the agkistrodon halys venom and application in the preparation of agkistrodon halys venom resistant serum for treating agkistrodon halys venom poisoning. According to the invention, the agkistrodon halys venom can be identified in a laboratory; agkistrodon halys venom is more accurately subjected toquality control, so that specific drug anti-agkistrodon halys poison serum for treating agkistrodon halys venom poisoning is produced; and the method is very important for ensuring the safety, effectiveness and quality controllability of anti-agkistrodon halys venom serum products and is of great significance for reference of the diagnosis and legal medical expert identification of the types of venom caused by snake venom poisoning. The method has the advantages of high detection sensitivity, high correct detection rate, no cross reaction and simple operation process.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and relates to a method for identifying snake venom using mass spectrometry, in particular to a method for identifying the type of snake venom using snake venom protein analysis technology; in addition, the invention also relates to the application of the identification method, identification The snake venom can be used in the production of antivenom, and has very important significance for the diagnosis of snake venom in snake poisoning and the reference of forensic identification. Background technique [0002] Venomous snakebites are common in subtropical and tropical regions. In the rural areas of Asia, Africa, Latin America and Oceania, venomous snakebite is a particularly prominent public health problem, and the incidence of snakebite in my country shows a trend of increasing year by year. [0003] The venom is mostly light yellow or milky white translucent viscous liquid with more than 10...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N27/447
CPCG01N27/447G01N30/02G01N30/06G01N30/72G01N2030/027G01N2030/062
Inventor 陈则杨智周东吴琼易应磊罗敏王德慧范铁炯
Owner SHANGHAI SERUM BIOTECH
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