Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Ochratoxin A hybridoma cell strain, antibody, compound immunoadsorbent, immunoaffinity column, kit and applications of immunoadsorbent, immunoaffinity column and kit

A hybridoma cell line and immunoadsorbent technology are applied in the field of immunoaffinity columns and kits to achieve stable performance.

Inactive Publication Date: 2014-07-02
INST OF CROP SCI CHINESE ACAD OF AGRI SCI +2
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The hybridoma cell line was deposited in the General Microorganism Center (CGMCC) of the China Committee for the Collection of Microorganisms on November 24, 2011

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
  • Ochratoxin A hybridoma cell strain, antibody, compound immunoadsorbent, immunoaffinity column, kit and applications of immunoadsorbent, immunoaffinity column and kit
  • Ochratoxin A hybridoma cell strain, antibody, compound immunoadsorbent, immunoaffinity column, kit and applications of immunoadsorbent, immunoaffinity column and kit
  • Ochratoxin A hybridoma cell strain, antibody, compound immunoadsorbent, immunoaffinity column, kit and applications of immunoadsorbent, immunoaffinity column and kit

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0029] Specifically, the preparation method of hybridoma cell line and monoclonal antibody in the present invention may comprise the following steps:

[0030] (1) Preparation of ochratoxin A immunogen

[0031] 2 mg of ochratoxin A (OTA) was added to 500 μL of acetone, and 4 mg of CDI was added to obtain a first solution. 8 mg of BSA was added to 3 mL of 0.1 M carbonate buffer at pH 9.6 to obtain a BSA solution. The first solution was slowly added dropwise to the BSA solution, and reacted at room temperature for 2 days. After the reaction, the solution was dialyzed against PBS solution for 3 days at 4°C.

[0032] (2) Preparation of hybridoma cells and monoclonal antibodies against ochratoxin A

[0033] Animal immunization: The immunized animals are female BALB / c mice about 6-8 weeks old. Five mice were immunized with the ochratoxin A immunogen. Take an appropriate amount of ochratoxin A immunogen (100 μg / cause) and add an equal amount of Freund's complete adjuvant to make a...

Embodiment 1

[0080] This example is used to prepare ochratoxin A immunogen and aflatoxin immunogen

[0081] (1) 2 mg of ochratoxin A (OTA) was added to 500 μL of acetone, and 4 mg of CDI was added to obtain the first solution. 8 mg of BSA was added to 3 mL of 0.1 M carbonate buffer at pH 9.6 to obtain a BSA solution. The first solution was slowly added dropwise to the BSA solution, and reacted at room temperature for 2 days. After the reaction, the solution was dialyzed against PBS solution for 3 days at 4°C. The ochratoxin A immunogen, OTA-BSA, was obtained.

[0082] (2) 4 mg of aflatoxin B 1 Dissolve in 2 mL of acetone, add 40 μL of 10% HO 2 SO 4 , Stir the reaction at 56°C for 4h; evaporate the reaction product to dryness, add 5mL H 2 O, extracted twice with 25 mL chloroform, then with 20 mL H 2 The organic layer was washed with O, and the organic layer was retained; the organic solvent was evaporated to obtain a yellow solid product.

[0083]Take 1.0 mg of the yellow solid produ...

Embodiment 2

[0085] This embodiment is used for preparing anti-aflatoxin B 1 , B 2 , G 1 , G 2 , M 1 , M 2 monoclonal antibody against ochratoxin A (antibody B)

[0086] (1) Animal immunization: the immunized animals were female BALB / c mice about 6-8 weeks old, and 5 mice were immunized with the ochratoxin A immunogen. The ochratoxin A immunogen (100 μg / cause) was added with an equal amount of Freund's complete adjuvant to make an emulsified agent for immunization, and then the adjuvant was changed to an incomplete adjuvant for 6 times of immunization, with an interval of 2 weeks between each time. Except for the first multi-point subcutaneous injection on the back of the neck, the rest were intraperitoneal injections. After the immunization, the mice were sacrificed, and splenocytes were collected.

[0087] Cell fusion: Splenocytes and hybridoma cells were subjected to cell fusion experiments at a ratio of 10:1.

[0088] Hybridoma cell cloning: The hybridoma cells were screened by...

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 hybridoma cell strain CGMCC NO. 5505 and a monoclonal antibody obtained by secretion of the cell strain, and an application of the antibody. The invention also provides an immunoadsorbent comprising a solid phase vector and antibodies coupled with the solid phase vector, wherein the antibodies are the above monoclonal antibody and a monoclonal antibody obtained by secretion of a cell strain CGMCC NO. 5506; and an immunoaffinity column loaded with the immunoadsorbent. The invention also provides a kit containing the above immunoadsorbent or the above immunoaffinity column, and applications of the above immunoadsorbent, the above immunoaffinity column and the kit in detecting aflatoxins, sterigmatocystins and ochratoxin A. Besides, the invention provides a separation method and a detection method specifically. The monoclonal antibody with stable and specific performance is developed by the invention, so that purification and detection for six aflatoxins, sterigmatocystins and ochratoxin A at the same time are realized.

Description

technical field [0001] The present invention relates to a hybridoma cell line, a monoclonal antibody secreted by the hybridoma cell line and its application, an immunoadsorbent prepared from the antibody, and an immunoaffinity column and kit equipped with the immunoadsorbent , and their role in the purification of aflatoxin B 1 , B 2 , G 1 , G 2 , M 1 , M 2 , versicolor and ochratoxin A application. Background technique [0002] Aflatoxins are a group of structurally similar secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus, etc., and are a group of compounds with difuranocoumarin as the basic structure. At present, 12 species have been isolated and identified, especially aflatoxin B 1 , B 2 , G 1 , G 2 , M 1 and M 2 It is a strong pollutant, widely present in grains, feed and its processed products. Among the more than 200 known mycotoxins, it is the most toxic and has the highest pollution frequency. Aflatoxins have the effects o...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/14G01N33/577C07K17/00C07K1/22C07K14/38
Inventor 李为喜王雄鲍蕾许艳丽王步军吕宁梁成珠果旗江帆王彦斐胡明勋邓颖戚大海吴兆广
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com