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Double-specificity monoclonal antibody resistant to triazophos and chlorpyrifos and preparation method and application thereof

A monoclonal antibody and anti-triazophos technology, applied in chemical instruments and methods, specific peptides, instruments, etc., can solve problems such as toxicity, single analysis technology and means, and achieve stable properties and high detection sensitivity

Inactive Publication Date: 2013-04-17
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Triazophos original drug is moderately toxic and toxic to fish and bees
[0010] Although there are currently reports on the immunoassay techniques for triazophos and chlorpyrifos, they are all limited to a single analysis technique and means. In the context of the need for multi-residue analysis, it is important to establish an immunoassay technique that can simultaneously detect two or two types of target compounds. It is particularly urgent, and the preparation of high-quality bispecific monoclonal antibodies is a new means and method for the establishment of immunological multi-residue analysis technology

Method used

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Examples

Experimental program
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Embodiment Construction

[0022] The present invention will be further described below in conjunction with the examples.

[0023] 1. Preparation of Immunogen

[0024] 1.1 Preparation of triazophos hapten, artificial antigen and coated antigen

[0025] 1.1.1 O - Synthesis of ethyl thiophosphoryl dichloride (TZLM-1)

[0026] Weigh the phosphorus trichloride (PSCl3 ) 68 g (about 0.4 mol) was placed in a three-necked flask with a low temperature thermometer, cooled to -10~-5 ℃ with an ice-salt water bath, and 55 g (about 1.2 mol) of absolute ethanol was added dropwise under vigorous stirring, Strictly control the rate of addition so that the temperature of the reaction solution is always not greater than 0 °C. After the dropwise addition, the reaction was continued at 10 °C for 2 h. After the reaction, the reaction solution (100 ml×2) was washed with (0±5)°C distilled water, the oil layer was separated and washed with anhydrous Na 2 SO 4 Dry it, then distill it under reduced pressure with a water pum...

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PUM

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Abstract

The invention discloses a double-specificity monoclonal antibody resistant to triazophos and chlorpyrifos and a preparation method and application thereof, and belongs to the technical field of biology. The double-specificity monoclonal antibody has specificity for triazophos and chlorpyrifos simultaneously. The preparation method for the monoclonal antibody comprises the steps as follows: synthesizing half antigens and artificial antigens of triazophos and chlorpyrifos; utilizing the artificial antigens of triazophos to prepare hybridoma cell strains secreting a component resistant to triazophos; utilizing the artificial antigens of chlorpyrifos to prepare hybridoma cell strains secreting a component resistant to chlorpyrifos; and after the triazophos-resistant hybridoma cell strains and hybridoma-resistant hybridoma cell strains are subjected to cell fusion, screening to obtain tetrasomic hybridoma cells which secrete the double-specificity monoclonal antibody resistant to triazophos and chlorpyrifos. The double-specificity monoclonal antibody prepared by the invention has the advantages that the characters are stable, the detection sensitivity is high, the double-specificity monoclonal antibody can be produced massively, and meanwhile the antibody is used for establishing an ELISA (enzyme-linked immuno sorbent assay) analysis method, so that the technical basis for the rapid detection of triazophos and chlorpyrifos is provided.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a bispecific monoclonal antibody against triazophos and chlorpyrifos, a preparation method and application thereof. Background technique [0002] Immunoassay (IA) is a technology that uses antibodies as biochemical detectors to perform qualitative and quantitative analysis of compounds, enzymes, or proteins. It is based on the specific and reversible binding reaction between antigens and antibodies. It has the characteristics of strong specificity, high sensitivity, convenience and speed, high throughput, low detection cost, safety and reliability. This method generally does not require expensive instruments, and the technical requirements of the users are not high. It is easy to popularize and promote, and is especially suitable for on-site screening and rapid analysis of a large number of samples. A series of detection products developed based on this technology, such ...

Claims

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Application Information

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IPC IPC(8): C07K16/44G01N33/577
Inventor 金仁耀朱国念桂文君
Owner ZHEJIANG GONGSHANG UNIVERSITY
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