Molecular modified penicillin-binding protein (PBP) capable of detecting beta-lactam antibiotic residues

A binding protein and molecular modification technology, applied in the field of rapid detection of β-lactam antibiotic residues, high affinity penicillin binding protein, can solve the problem of insufficient affinity

Inactive Publication Date: 2014-06-18
JILIN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the known affinity between penicillin-binding proteins and β-lactam antibiotics does not meet the detection standards required by the state. In order to achieve rapid detection of β-lactam antibiotics, it is urgent to find a protein with higher affinity for β-lactam antibiotics. penicillin binding protein

Method used

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  • Molecular modified penicillin-binding protein (PBP) capable of detecting beta-lactam antibiotic residues
  • Molecular modified penicillin-binding protein (PBP) capable of detecting beta-lactam antibiotic residues
  • Molecular modified penicillin-binding protein (PBP) capable of detecting beta-lactam antibiotic residues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0021] Example colloidal gold test strip assembly

[0022] A beta-lactam antibiotic rapid detection test strip includes: a bottom plate, a sample pad, a colloidal gold pad, a nitrocellulose membrane, an absorption pad, a detection line, and a quality inspection line (see Figure 3). The colloidal gold pad has colloidal gold labeled JN-Pro-β-lactam, the nitrocellulose membrane is equipped with a detection line and a quality inspection line, the detection line has ampicillin coupled antigen, and the quality control line has JN-Pro-β -Monoclonal antibody to lactam.

[0023] A method for making test strips for rapid detection of beta-lactam antibiotics includes:

[0024] (1) Preparation of JN-Pro-β-lactam

[0025] For details of the preparation process, see "Specific Implementation Mode".

[0026] (2) Preparation of coupling antigen of bovine serum albumin and ampicillin

[0027] 25mg ampicillin dissolved in pH9.6Na 2 CO 3 Add 5mg BSA to the buffer solution, adjust the pH to 11, stir at 37°...

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Abstract

The invention provides a molecular modified penicillin-binding protein (PBP) capable of detecting beta-lactam antibiotic residues. The molecular modified PBP is characterized by being obtained by respectively modifying leucine on the 147th position of the PBP of actinomadura yumaense R39 and histidine on the 351st position of the PBP of the actinomadura yumaense R39 into histidine and aspartic acid, so that the affinity of beta-lactam antibiotics is improved. The invention also provides a rapid detecting test strip based on production of the protein, and the rapid detecting test strip can be used for rapidly detecting the beta-lactam antibiotic residues of agricultural and sideline products.

Description

Technical field [0001] The invention relates to the field of food safety detection. More specifically, the present invention relates to a high-affinity penicillin binding protein obtained by molecular transformation and its application in rapid detection of β-lactam antibiotic residues in agricultural and sideline products. Background technique [0002] Dairy cows are susceptible to mastitis, vaginitis and other diseases during the breeding process. Due to the good efficacy and low price of β-lactam antibiotics, dairy farmers use a large number of them when dairy cows are sick, resulting in residual β-lactam antibiotics in raw milk The phenomenon is widespread, which has greatly increased the difficulty of milk source quality and safety control for dairy products companies, and dairy products containing β-lactam antibiotic residues will affect the health and life safety of consumers. Therefore, the Ministry of Agriculture has issued the industry standard "Non-polluted Food Fresh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/36C12N1/21G01N33/68C12R1/19
CPCC07K14/195G01N33/68
Inventor 李铁柱张杰李倬林曾宪鹏
Owner JILIN ACAD OF AGRI SCI
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