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Multiple cluster antigen and its preparation, wide spectrum specific multiple clone antigen and its use

A polyclonal antibody and multi-cluster antigen technology, which is applied in the field of multi-cluster antigen and preparation, polyclonal antibody, can solve the problems of increasing the recognition range, application limitations, and the uncertainty of the antibody recognition range, so as to save money and improve work efficiency. Effect

Inactive Publication Date: 2007-02-07
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

While increasing the recognition range of this method, the detection sensitivity decreases, and the recognition range of this type of antibody cannot be determined in advance
In addition, the application of this method in the multi-residue detection of compounds of different structural types is limited, and there has been no relevant literature report that an antibody can simultaneously recognize a variety of small molecule compounds of different structural classes

Method used

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  • Multiple cluster antigen and its preparation, wide spectrum specific multiple clone antigen and its use
  • Multiple cluster antigen and its preparation, wide spectrum specific multiple clone antigen and its use
  • Multiple cluster antigen and its preparation, wide spectrum specific multiple clone antigen and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The haptens used are ethyl parathion (PA) and fenvalerate hapten (JU), the structural formula is as follows:

[0024]

[0025] Preparation of immunogen JU-BSA-PA

[0026] (1) The synthesis of the immunogen uses the carbodiimide method: 80 micromoles of the organic phosphorus pesticide ethyl parathion hapten compound PA is dissolved in 2 ml of N, N-dimethylformamide, and the equivalent is added Dicyclohexylcarbodiimide and N-hydroxysuccinimide, react at room temperature, centrifuge, take 100~800μL of supernatant and slowly add to 4~8mL10~20mg / mL bovine serum albumin (BSA) carbonate In the buffer solution, react for 1 to 6 hours under magnetic stirring, put it into a dialysis bag, first dialyzed with distilled water for 2 to 4 times, then dialyzed with normal saline, and store at -20°C for later use.

[0027] (2) Using the product obtained in the above steps as a carrier, use the same method to continue to couple another small molecule compound hapten molecule JU, and then ...

Embodiment 2

[0050] The haptens used are ethyl parathion (PA) and carbofuran hapten (BFNB). The structural formula is as follows:

[0051]

[0052] Preparation of immunogen PA-BSA-BFNB

[0053] (1) The synthesis of immunogen uses the carbodiimide method: dissolve 80 micromoles of carbamate pesticide carbofuran hapten compound BFNB in ​​2ml of N,N-dimethylformamide, and add the equivalent amount Dicyclohexylcarbodiimide and N-hydroxysuccinimide, react at room temperature, centrifuge, take 100~800μL of supernatant and slowly add to 4~8ml carbonate buffer containing 10~20mg / mL BFNB-BSA In the solution, react under magnetic stirring for 1 to 6 hours, put it into a dialysis bag, first dialyzed with distilled water for 2 to 4 times, then dialyzed with saline, and store at -20°C for later use.

[0054] (2) Using the product obtained in the above steps as a carrier, use the same method to continue to couple another small molecule compound hapten molecule BFNB, and then perform dialysis purification;...

Embodiment 3

[0077] The haptens used are triazophos hapten (THBu), chlorpyrifos hapten (DSP) and carbofuran hapten (BFNB), the structural formula is as follows:

[0078]

[0079] Immunogen Preparation

[0080] (1) Preparation of BSA-DSP conjugate: dissolve 80 micromoles of the hapten compound DSP of the organophosphorus pesticide chlorpyrifos in 2ml of N,N-dimethylformamide, and add an equivalent amount of dicyclohexylcarbodiimide Amine and N-hydroxysuccinimide, react at room temperature, centrifuge, take 100~800μL of supernatant and slowly add 4~8mL of 10~20mg / mL bovine serum albumin (BSA) carbonate buffer solution, stir under magnetic force React for 1 to 6 hours, put it into a dialysis bag, first dialyzed with distilled water for 2 to 4 times, then dialyzed with saline, and store at -20°C for later use.

[0081] (2) Using the product obtained in the above steps as a carrier, use the same method to continue coupling another small molecule compound hapten molecule BFNB, and then perform di...

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Abstract

The disclosed preparation method for multi-cluster antigen comprises: selecting 2-15 different micro-molecular compound semi-antigens, and 1) coupling one micro-molecular semi-antigen to the protein molecule for dialysis and purification; 2) with last product as carrier, continuing to couple another micro-molecular semi-antigen for further dialysis and purification; 3) repeating step 2) till coupling all the semi-antigens. This invention has specificity of broad spectrum.

Description

Technical field [0001] The invention belongs to the technical field of immunochemistry and residue analysis of small molecule compounds (molecular weight less than 1000 Daltons), and specifically relates to a multi-cluster antigen and a preparation method, as well as a polyclonal antibody and its use. Background technique [0002] The antibodies currently used for immunochemical analysis are mainly monoclonal antibodies and polyclonal antibodies, most of which are for the detection of single target compounds, with high specificity and specificity. But for multi-residue analysis of small molecule compounds such as pesticides and veterinary drugs, immunological methods usually have two approaches. One is to first prepare corresponding specific antibodies for different pesticides and other small molecule compounds, and then use these antibodies together. It can detect multiple pesticides and other small molecule compounds simultaneously. Another method is to design and synthesize ha...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N33/53
Inventor 桂文君朱国念王姝婷郭逸蓉金仁耀
Owner ZHEJIANG UNIV
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