Unlock instant, AI-driven research and patent intelligence for your innovation.

Anti-methyl mercury monoclonal antibody hybridoma cell strain GZ02 and application thereof

A hybridoma cell line, anti-methylmercury single technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, instruments, etc., can solve the problems of difficult on-site detection, long time-consuming detection, high cost, etc., and achieve good detection sensitivity and specificity, good stability, and high antibody specificity

Active Publication Date: 2017-07-14
JIANGNAN UNIV +1
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection of methylmercury mostly adopts high-performance liquid chromatography-fluorescence spectroscopy, high-performance liquid chromatography-plasma chromatography and other instrument detection methods, which are time-consuming and expensive, and are difficult to be used for on-site detection.

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
  • Anti-methyl mercury monoclonal antibody hybridoma cell strain GZ02 and application thereof
  • Anti-methyl mercury monoclonal antibody hybridoma cell strain GZ02 and application thereof
  • Anti-methyl mercury monoclonal antibody hybridoma cell strain GZ02 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Preparation of hybridoma cell line GZ02

[0021] (1) Synthesis of complete antigen: Dissolve 4.5mg MNA, 30mg DCC, and 16.7mg NHS in 1mL DMF, stir overnight at room temperature; centrifuge at 10,000rpm for 10min, discard the precipitate, and keep the supernatant. Take 15mgBSA and dissolve it in 0.13mol / L NaHCO 3 in solution. The above supernatant was added dropwise to the BSA solution, and stirred at room temperature for 8 h. Centrifuge at 10,000 rpm for 10 min, and dialyze the supernatant in 0.01 mol / L PBS solution for 3 days to prepare MNA-BSA solution. Take 2mg CH 3 HgCl was dissolved in 1 mL methanol / NaOH (1M) (V / V9:1) solution, then added dropwise to the MNA-BSA solution, stirred at room temperature overnight, and then dialyzed in 0.01 mol / L PBS solution for 2 days, Produced CH 3 Hg-MNA-BSA, as an immune antigen, was stored at -20°C. Coated original CH 3 The synthesis method of Hg-MNA-OVA is the same as above.

[0022] (2) Animal immunization: hea...

Embodiment 2

[0031] Example 2 Application of anti-methylmercury specific monoclonal antibody

[0032]The monoclonal antibody prepared by hybridoma cell line GZ02 through in vivo ascites was applied to the indirect competitive ELISA detection of methylmercury in water, and the specific steps were as follows:

[0033] (1) Encapsulation: the original encapsulation (CH 3 Hg-MNA-OVA) was diluted to 1.0 μg / mL with 0.05M pH9.6 carbonate buffer, 100 μL / well was added to the microtiter plate, and reacted at 37°C for 2 hours;

[0034] (2) Washing: Pour off the solution in the plate, spin dry, and wash 3 times with washing liquid, 3 minutes each time;

[0035] (3) Blocking: After patting dry, add 200 μL / well blocking solution and react at 37°C for 2 hours. After washing, dry it for later use;

[0036] (4) Add sample: add CH 3 HgCl and antibody: dilute CH with 0.01M PBS buffer 3 Add 50 μL of HgCl to each well, then add 50 μL of antibody solution diluted by a certain factor, and react at 37°C for ...

Embodiment 3

[0047] Hybridoma cell line GZ02 monoclonal antibody against CH 3 Cross-reactivity rates of HgCl, Cd(II), Cu(II), Ca(II), Zn(II), Mn(II), Mg(II), Pb(II). As shown in Table 1.

[0048] Table 1 Hybridoma cell line GZ02 monoclonal antibody to CH 3 Cross-reactivity rates of HgCl, Cd(II), Cu(II), Ca(II), Zn(II), Mn(II), Mg(II), Pb(II)

[0049]

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

PropertyMeasurementUnit
Sensitivityaaaaaaaaaa
Login to View More

Abstract

An anti-methyl mercury monoclonal antibody hybridoma cell strain GZ02 and an application thereof. The invention belongs to the technical field of food safety detection. The hybridoma cell strain GZ02, which secretes an anti-methyl mercury monoclonal antibody, is prepared by: mixing and emulsifying a methyl mercury complete antigen with a Freund's adjuvant in equal amount, immunizing BALB / c mice through back subcutaneous injection, and performing indirect competitive ELISA screening and three-time subcloning. The hybridoma cell strain has been preserved in China General Microbiological Culture Collection Center and is assigned the accession number of CGMCC No.13081. The secreted mono-antibody can recognize the methyl mercury with high specificity and sensitivity, and can be used for detection of residue of the methyl mercury in food safety. The antibody has high detection sensitivity and specificity to methyl mercury, IC50 value being 23 ng / mL. The antibody also has good stability. The hybridoma cell strain GZ02 can be preserved for at least 5 years by cryopreservation in liquid nitrogen, and purified antibody can be preserved for at least 2 years in a refrigerator at -20 DEG C. The antibody has high specificity and no cross reaction is found between the antibody and other metal ions so far.

Description

technical field [0001] The invention discloses an anti-methylmercury monoclonal antibody hybridoma cell line GZ02 and its application, relates to the hybridoma cell line GZ02 and the anti-methylmercury monoclonal antibody secreted by the cell line, and belongs to the technical field of food safety detection. Background technique [0002] Mercury (mercury, Hg) and its compounds are widely used in industrial and agricultural fields, but it is also a highly toxic heavy metal pollutant. At present, mercury pollution in the environment is becoming more and more serious. Experiments have shown that no matter for elemental mercury, inorganic mercury, or organic mercury, they can directly enter the human body through ingestion, skin, breathing and other channels or indirectly accumulate in animals and humans through food chain enrichment, resulting in multiple organs, multiple The system is poisoned. In general, organic mercury is more toxic than inorganic mercury. Among them, ino...

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 Applications(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/577C12R1/91
CPCC07K16/44G01N33/577G01N2430/00
Inventor 胥传来邹淑珍匡华徐丽广马伟刘丽强吴晓玲宋珊珊冯民
Owner JIANGNAN UNIV
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com