Snake toxin antigen-antibody reaction quantitative determination method

A technology for antibody reaction and quantitative determination, applied in the field of biochemistry, can solve the problems of strong carcinogenicity, low separation rate, loss of antigen, etc., and achieve the effects of simple operation, high resolution, and good application prospects.

Inactive Publication Date: 2007-10-24
广州医学院
View PDF2 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Western blotting still has certain deficiencies in the practical application of snake venom and anti-venom antibody quality inspection, such as the inevitable loss of some antigens during

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Snake toxin antigen-antibody reaction quantitative determination method
  • Snake toxin antigen-antibody reaction quantitative determination method
  • Snake toxin antigen-antibody reaction quantitative determination method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Determination of the Optimal Ratio of Antigen / Antibody Binding

[0042] (1) Cobra venom was extracted with a snake venom extractor (patent number: ZL200420046279.9), prepared into standard cobra venom freeze-dried powder, and stored at 4°C for later use.

[0043](2) After emulsifying the standard snake venom antigen and Freund's adjuvant, inject it into the chicken body, collect anti-venom IgY eggs after 2 weeks, use water dilution method to extract and ammonium sulfate salting out-anion exchange chromatography and other steps (refer to the patent application number : Publication of 200610035761.6), to prepare the monovalent anti-venom chicken egg yolk antibody (IgY) of cobra.

[0044] (3) Take the standard cobra venom freeze-dried powder and anti-cobra venom IgY to prepare standard cobra venom solution and standard anti-cobra venom IgY solution respectively. The standard cobra venom was mixed with the standard anti-cobra venom IgY in different proportions (...

Embodiment 2

[0050] Embodiment two: anti-cobra venom IgY stability determination

[0051] The extraction and preparation of the standard cobra venom and the standard anti-cobra venom IgY were the same as in Example 1. After the cobra venom solution and the anti-cobra venom IgY solution were obtained, the acid-base stability and thermal stability of the anti-cobra venom IgY were determined.

[0052] (1) Determination of acid-base stability of anti-cobra venom IgY

[0053] Use 1mol / l HCL or NaOH to adjust the pH value of the standard anti-cobra venom IgY solution to 2, 3, 4, 5, 6, 6.5, 7, 8, 9, 10, 11 and 12, and let it stand at room temperature for 30 minutes Mix with standard cobra venom at 2.75:1, incubate at 38°C for 45 minutes, shake for 15 minutes at a speed of 100 rpm / min; let stand for 30 minutes. Then add sample buffer, so that the final concentration of standard cobra venom is 10ug / ul, and the final concentration of anti-cobra venom IgY is 27.5ug / ul. React at 100°C for 5 minutes,...

Embodiment 3

[0064] Example 3: Determination of cross-immunity activity

[0065] Take the standard anti-cobra venom IgY and mix them with Chinese cobra, Agkistrodon akistrodon, domestic viper and Bungara venom at 2.75:1 respectively. Add 10ul. Stained after electrophoresis, the gel was scanned by optical density, and the cross-immune activity and potency between monovalent antivenom IgY and four common venomous snake venoms in my country were quantitatively analyzed according to the disappearance or density reduction of the relevant snake venom bands. The result shows (referring to Fig. 4): compare with swimming lane 1, all bands of cobra venom in swimming lane 2 all drift, illustrate that anti-cobra venom can completely neutralize cobra venom; Compared with swimming lane 7, in swimming lane 8, only part The band drifted or the density decreased, indicating that the anti-cobra venom could partially neutralize the silver ring snake venom; after the co-incubation of viper venom and Agkistro...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for quantitatively testing venom antigen-antibody reaction, comprising that (1), preparing venom antigen and antibody, (2), mixing the antigen and antibody to be arranged into a constant-temperature vibrator to be pre-cultivated, as first vibration and later laid stably, (3), adding a sample buffer liquid into the antigen-antibody solution, for 5-15min at 100Deg. C, eccentrically treating while the obtained upper clear liquid is the sample liquid, (4), using SDS-polyacrylamide gel electrophoresis to process electrophoresis and development on the sample solution, according to the density, change and disappear condition of the color band of electrophoresis channel, quantitatively analyzing the antigen-antibody reaction character. The invention quantitatively analyzes the purity, molecule weight and immunity character of the venom antigen and antibody, and test the optimized ratio of combined antigen and antibody, to prepare venom drug, which can save antibody and improve treatment effect.

Description

technical field [0001] The invention relates to a method for quantitative determination of antigen-antibody reaction in the field of biochemistry, in particular to an SDS-polyacrylamide gel electrophoresis (SDS-PAGE) method for quantitative determination of snake venom antigen-antibody reaction. Background technique [0002] The venom (snake venom) of vipers is a huge treasure house of biological resources. Since British Reid discovered for the first time in 1963 that the Malaysian red-mouthed Agkistrodon contained thrombin-like enzymes that caused the decline of fibrinogen, the comprehensive utilization and development of snake venom has been paid more and more attention by people. In 1968, the British successfully developed the snake venom preparation ARIN and first put it on the market to treat vascular occlusive diseases. In recent years, with dozens of snake venom products (drugs) coming out, and achieved good clinical curative effect and sales performance, the develop...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N33/559
Inventor 孔天翰董伟华刘四红祁俊华陈熹陈学文
Owner 广州医学院
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap