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Cell strain 16, monoclonal antibody produced by cell strain 16 and use of monoclonal antibody produced by cell strain 16

A monoclonal antibody and cell line technology, applied in the field of cell lines, can solve the problems of high cost, cumbersome process, and many hapten synthesis steps, and achieve the effects of good market prospects, strong specificity and high sensitivity

Active Publication Date: 2014-07-09
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of producing antibodies, the hapten design and synthesis are first carried out. The hapten synthesis of lambda-cyhalothrin has many steps, the process is cumbersome, and contains a variety of optical isomers. Therefore, the hapten of lambda-cyhalothrin Antibody preparation is difficult
So far, there are only reports on the preparation of polyclonal antibodies to cyhalothrin [1]; compared with polyclonal antibodies, the preparation process of monoclonal antibodies is long and costly, but it is easy to reproduce, with relatively stable quality and specificity. Strong, so more popular in the market
However, so far, there is no report on the preparation of monoclonal antibodies to lambda-cyhalothrin

Method used

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  • Cell strain 16, monoclonal antibody produced by cell strain 16 and use of monoclonal antibody produced by cell strain 16
  • Cell strain 16, monoclonal antibody produced by cell strain 16 and use of monoclonal antibody produced by cell strain 16
  • Cell strain 16, monoclonal antibody produced by cell strain 16 and use of monoclonal antibody produced by cell strain 16

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Implementation example 1 Application of enzyme-linked immunoassay based on anti-lambda-cyhalothrin monoclonal antibody to detection of lambda-cyhalothrin residues in water samples

[0048] 1. Determination of sensitivity of monoclonal antibody against lambda-cyhalothrin

[0049] Indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was used to measure the inhibitory effect of monoclonal antibodies on lambda-cyhalothrin. The concentration of the original coating H2-BSA was 0.5 mg / mL, and the protein concentration of the antibody was 0.06 mg / mL. , the measurement results were fitted with four-parameter regression using origin 8.5 software, the results are shown in Figure 4 . According to the fitted regression equation, the calculated antibody IC50 was 13.26±1.23ng / mL; the linear range of the standard curve was between 1-100 ng / mL; the detection limit reached 1.83ng / mL.

[0050] 2. Sample pretreatment and measurement Taihu Lake water was selected as the wa...

Embodiment 2

[0051] Implementation Example 2 Application of colloidal gold immunochromatographic test strips based on anti-lambda-cyhalothrin monoclonal antibody for the detection of lambda-cyhalothrin residues in fruits and vegetables

[0052] 1. Determination of sensitivity of monoclonal antibody against lambda-cyhalothrin

[0053] Prepare a series of standard solutions of lambda-cyhalothrin: 0 ng / mL, 10 ng / mL, 20 ng / mL, 50 ng / mL, 100 ng / mL, 200 ng / mL, 500 ng / mL, 1000 ng / mL; Insert the sample pad of the test strip into the standard solution of lambda-cyhalothrin, the solution flows through the test strip through chromatography, and after 10-12 minutes, the determination result of the gold standard test strip See Figure 4 , the naked-eye sensitivity of colloidal gold immunochromatographic test strips for detecting lambda-cyhalothrin reached 200 ng / mL; it can be seen that the monoclonal antibody secreted by cell line 16 can be used for the detection of lambda-cyhalothrin pesticide res...

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Abstract

The invention relates to a cell strain 16, a monoclonal antibody produced by the cell strain 16 and a use of the monoclonal antibody produced by the cell strain 16. A preparation method of the cell strain 16 comprises the following steps of coupling hapten 1 (H1) 3-[cyano[cis-3-[2-chloro-3,3,3-trifluoroethyl-2, dimethyl]cyclopropane-phenoxy]phenylpropionic acid and hemocyanin KLH by an active ester method, coupling hapten 2 (H2) cyano-[3-(4-aminophenoxy)phenyl-cis-3-[2-chloro-3,3,3-trifluorovinyl-2,2-dimethyl]cyclopropionate and bovine serum albumin by a diazotization method to obtain a conjugate as a coating antigen, and carrying out animal immunization, serum determination, cell fusion, screening and subcloning to obtain the cell strain 16. Through a mouse internal ascites induced-production method, an ascitic fluid is produced, and then through a caprylic acid-ammonium sulfate precipitation method, the ascitic fluid is purified so that the corresponding monoclonal antibody is produced. The monoclonal antibody has high sensitivity, median inhibitory concentration (IC50) of 13.26+ / -1.23ng / mL and strong singularity, provides a technology for detection of high-efficiency cyhalothrin residue in domestic agricultural products and has a good market prospect.

Description

technical field [0001] The invention relates to a cell line, specifically a cell line No. 16, an anti-effective-cyhalothrin monoclonal antibody produced by the cell line, and an application thereof. Background technique [0002] Beta-cyhalothrin, also known as cyhalothrin (CAS No. 91465-08-6), chemical name: (IR / IS)-cis-(Z)-2,2-dimethyl-3-( 2-Chloro-3,3,3-trifluoro-1-propenyl)cyclopropanecarboxylic acid-(S)-a-cyano-3-phenoxybenzyl ester (1:1 mixture), is a Pyrethroid insecticides and acaricides have the functions of avoiding, knocking down and poisoning. They have a wide insecticidal spectrum, high activity, rapid drug effect, and are resistant to rain after spraying. They are widely used in fruit trees, vegetables, cotton, and tobacco. , corn and other crops. With the widespread application of lambda-cyhalothrin, it has caused some impact on the ecological environment. Toxicological studies have shown that lambda-cyhalothrin is highly toxic to fish and aquatic arthropod...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/577C12R1/91
Inventor 陈秀金胥传来郭金英任国艳李道敏孙军杰张敏辛莉
Owner HENAN UNIV OF SCI & TECH
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