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Polypeptide capable of specifically binding to benzothiostrobin immune complex and application thereof

A technology of immune complex and benthiazolin, which can be used in peptides, material testing products, biological testing, etc., can solve problems such as peptides that have not yet been seen, and achieve the effect of high accuracy, strong specificity and high sensitivity

Active Publication Date: 2015-12-16
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the non-competitive assay mode has advantages, the technology is still in the early stage of research and development, and the biggest challenge in establishing this assay is the screening of anti-immune complex peptides
So far, there have been no studies and reports on peptides that specifically bind to benzathystrobin immune complexes and their applications

Method used

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  • Polypeptide capable of specifically binding to benzothiostrobin immune complex and application thereof
  • Polypeptide capable of specifically binding to benzothiostrobin immune complex and application thereof
  • Polypeptide capable of specifically binding to benzothiostrobin immune complex and application thereof

Examples

Experimental program
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Embodiment 1

[0028] Example 1 Panning and preparation of benzathystrobin immune complex specific binding polypeptide

[0029] 1. Panning of phage clones that specifically bind to benzathystrobin immune complex

[0030] According to the cycle of "adsorption-washing-elution-amplification", after three rounds of panning, the specific operation is as follows:

[0031] (1) Take 100 μL of 10 μg / mL protein A column-purified penthiostrobin antibody and add it to the microtiter plate, and coat at 4°C overnight, with a total of three wells;

[0032] (2) Take a small amount of Escherichia coli ER2738 and spread it on the LB+Tet plate, culture overnight at 37°C;

[0033] (3) Wash the ELISA plate in step (1) 5 times with PBST, add 300 μL of 3% BSA, incubate at 37° C. for 1 hour, wash 5 times with PBST, and store at 4° C. for later use;

[0034] (4) Add 10 μg / mL of penthiostrobin to the microtiter plate in step (3), shake slightly at room temperature for 1 hour, and wash 5 times with PBST;

[0035] (...

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Abstract

The invention belongs to the technical field of biology, relates to a polypeptide capable of specifically binding to a benzothiostrobin immune complex, and comprises application of the polypeptide in detection of benzothiostrobin. According to the invention, with a phage display technology, a phage display random cycle octapeptide library undergoes 3 rounds of panning by using the benzothiostrobin immune complex, and phage clones capable of binding to the benzothiostrobin immune complex are panned out; and a plurality of phage clones are randomly selected for amplification and plasmid extraction, and positive phage clones are selected through a non-competitive phage ELISA method and are sequenced so as to obtain polypeptide sequences. The invention also relates to the application of the phage display polypeptide in the detection of the benzothiostrobin. The non-competitive phage ELISA method established by using the phage display polypeptide can be applied in rapid, sensitive, simple, convenient and cheap detection of benzothiostrobin residues in the environment and agricultural products.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a polypeptide that specifically binds to a penthiostrobin immune complex, including the application of the polypeptide in detecting penthiostrobin. Background technique [0002] Benzothiostrobin is a new type of Strobilurins fungicide developed by Central China Normal University. Penthiostrobin has the advantages of wide bactericidal spectrum, high bactericidal activity, low application rate, long-lasting effect, etc. It has protective and therapeutic effects, and has excellent control effects on crop powdery mildew and downy mildew, and has a good market Development prospects. In order to prevent the potential risks in the application of penthiostrobin, a sensitive, rapid and selective residue detection method is needed. [0003] At present, the residue detection of fenthiastrobin includes instrumental analysis method and immunoassay method. The immunoassay method has the advantage...

Claims

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Application Information

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IPC IPC(8): C07K7/06G01N33/68
Inventor 华修德王鸣华周亮亮丁园
Owner NANJING AGRICULTURAL UNIVERSITY
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