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Preparation method of peptide ligand for non-competitive detection of water-soluble pesticide residues

A fat-soluble pesticide, non-competitive technology, applied in the direction of peptides, etc., can solve the problems of difficult to achieve high-capacity requirements, cumbersome process of antibody library construction, etc., to achieve the effect of shortening the preparation cycle, simple and effective operation, and improving screening efficiency.

Active Publication Date: 2015-06-10
HUNAN UNIV OF ARTS & SCI +1
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  • Claims
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Problems solved by technology

In the process of preparing antibodies using phage display technology, it is usually necessary to construct a high-capacity phage display antibody library first. However, the process of antibody library construction is usually cumbersome, and the high-capacity requirement is usually difficult to meet

Method used

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  • Preparation method of peptide ligand for non-competitive detection of water-soluble pesticide residues
  • Preparation method of peptide ligand for non-competitive detection of water-soluble pesticide residues
  • Preparation method of peptide ligand for non-competitive detection of water-soluble pesticide residues

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Embodiment Construction

[0039] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0040] Chlorpyrifos is an organophosphorus pesticide widely used at present, and its residue problem has also been paid attention to. In the following examples, the inventors provide a method for preparing a peptide ligand for the non-competitive detection of chlorpyrifos residues. On the basis of preparing β-CD-BSA coupling products, the method uses cyclodextrins to chlorpyrifos. Inclusion effect, the peptide ligand that can specifically recognize the "β-CD-chlorpyrifos" complex was screened from a commercial phage display peptide library using the "positive and negative alternate screening" strategy, and its amino acid sequence was obtained.

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Abstract

The invention discloses a preparation method of a peptide ligand for non-competitive detection of water-soluble pesticide residues. The preparation method comprises the following steps: coupling beta-cyclodextrin (beta-CD) with bovine serum albumin (BSA), enveloping the coupling products with a solid phase carrier to complex with chlorpyrifos; performing four-round screening on commercialized phage display peptide libraries by using 'positive and negative alternative screening' strategy; separating and selecting mono phage clones from the fourth-round screening elution products, identifying the mono phage clone capable of specifically recognizing 'beta-CD-pesticide' complex; finally sequencing the identified phage clone to determine the insertion of polypeptide fragment and obtain the amino acid sequence information of the polypeptide. The preparation method has the advantages that the molecular modification of the detection object is not needed, and the obtained peptide ligand is used for constructing the non-competitive detection of water-soluble pesticide residues.

Description

technical field [0001] The invention relates to a method for preparing a peptide ligand for non-competitive detection of fat-soluble pesticide residues, belonging to the field of biotechnology and analysis and detection technology. Background technique [0002] The problem of pesticide residues is one of the food safety and environmental safety issues that are widely concerned. The premise of its control is to establish an effective detection technology system. The use of rapid detection methods to conduct preliminary screening of possible pesticide residues in food or the environment is recognized by domestic and foreign counterparts as an important part of the pesticide residue detection system. Although many studies have been carried out on the immunological rapid detection of pesticide residues, there are not many examples of its practical application so far. The cumbersome antibody preparation process and the single analysis and detection mode are the main bottleneck f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08
Inventor 贺江
Owner HUNAN UNIV OF ARTS & SCI
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