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PBP-1A affinity beta-lactam antibiotic electrochemical biosensor, and making method and application thereof

A PBP-1A, biosensor technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of difficulty in obtaining penicillin antibodies, influence of detection efficiency, and no disclosure of antibiotic electrochemical biosensors, etc., to reduce the detection limit. , low detection limit, the effect of improving analytical sensitivity

Active Publication Date: 2015-03-11
NINGBO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the limitations of detection methods and conditions, the above-mentioned standards have not been strictly implemented, resulting in the problem of penicillin residues in animal products in my country has not been resolved, which objectively requires the improvement of traditional detection methods, the introduction of advanced detection instruments, and the search for A fast and accurate detection method, making the detection results more accurate and reliable
[0005] In 2012, Babington published the article "Current Status of Bioanalytical Methods for Penicillin Detection" in "Analytical and Bioanalytical Chemistry". The detection efficiency of ELISA is also greatly affected
[0006] However, at present, there are no relevant research reports on the preparation of PBP-1A affinity β-lactam antibiotic electrochemical biosensors by combining graphene, PBP, and electrochemical analysis methods at home and abroad.

Method used

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  • PBP-1A affinity beta-lactam antibiotic electrochemical biosensor, and making method and application thereof
  • PBP-1A affinity beta-lactam antibiotic electrochemical biosensor, and making method and application thereof
  • PBP-1A affinity beta-lactam antibiotic electrochemical biosensor, and making method and application thereof

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specific Embodiment 1

[0039] A PBP-1A affinity type β-lactam antibiotic electrochemical biosensor, the glassy carbon electrode is the working electrode, and the surface of the working electrode is modified with the nanomaterial graphene, the recognition element penicillin G and the binder nafion, as As for the detection carrier, the above-mentioned PBP-1A is penicillin-binding protein-1A derived from Streptococcus pneumoniae.

specific Embodiment 2

[0040] The preparation method of the PBP-1A affinity type β-lactam antibiotic electrochemical biosensor of the above-mentioned embodiment 1, the experimental materials are glassy carbon electrodes, Al 2 o 3 The powder, suede and glass plate, iron stand, and electrochemical analyzer were purchased from Shanghai Chenhua Instrument Co., Ltd., absolute ethanol, Na 2 HPO 4 , NaH 2 PO 4 etc. are analytically pure from Shanghai Sinopharm. Deionized water was prepared in the laboratory. Specifically include the following steps:

[0041] (1) Polishing and cleaning of the bare electrode: the glassy carbon electrode is sequentially cleaned with Al 2 o 3 The powder was polished into a mirror surface, and then ultrasonically cleaned with absolute ethanol and deionized water for 5 minutes respectively; after the peak voltage difference was detected by cyclic voltammetry to be less than 80mV, the electrode surface was rinsed with PBS solution with a pH of 7.4, and dried;

[0042] (2...

specific Embodiment 3

[0052] The sensitivity and linear range of the PBP-1A affinity type β-lactam antibiotic electrochemical biosensor to β-lactam antibiotics, the specific steps are as follows:

[0053] (1) Polishing and cleaning of the bare electrode: the glassy carbon electrode is sequentially cleaned with Al 2 o 3 The powder was ground into a mirror surface, and then ultrasonically cleaned with absolute ethanol and deionized water for 5 minutes respectively (ultrasonic cleaner, Kunshan Shumei KQ3200E); an electrochemical analyzer (Shanghai Chenhua CHI852c) was used for cyclic voltammetry (CV method) ) After detecting the peak voltage difference 2 blow dry;

[0054] (2) Activation of the bare electrode: place the glassy carbon electrode obtained in step (1) in a 1mol / L sulfuric acid solution, and scan for 10 cycles by CV method until it is stable;

[0055] (3) Modification of the bare electrode: Add 4 μL of 2.5 mg / mL nanomaterial graphene and 2 μL of 0.2 g / L recognition element penicillin G...

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Abstract

The invention discloses a PBP-1A affinity beta-lactam antibiotic electrochemical biological sensor, and a making method and an application thereof. The electrochemical biological sensor is characterized in that the surface of a glassy carbon electrode is commonly modified by graphene, penicillin G and nafion. The making method comprises the steps of bare electrode polishing, cleaning and activating, and concretely comprises: sequentially adding graphene and penicillin G to the surface of the glassy carbon electrode in a dropwise manner, well mixing, carrying out blow drying at 37DEG C, adding nafion in a dropwise manner, naturally air-drying the surface, immersing the electrode in 5wt% bovine serum albumin, placing the immersed electrode in an oven, drying at 37DEG C for 30min, taking out the electrode, washing the electrode with a PBS solution to obtain a glassy carbon electrode with the surface modified by graphene-penicillin G-nafion as a work electrode, placing the work electrode in a system solution, carrying out DPV detection, and determining the concentration of beta-lactam antibiotics in a sample to be detected according to a standard curve. The electrochemical biological sensor has the advantages of high sensitivity, fast detection speed, low detection limit, high efficiency and cheapness.

Description

technical field [0001] The invention relates to the field of food safety detection, in particular to a PBP-1A affinity β-lactam antibiotic electrochemical biosensor for detecting antibiotic residues in dairy products, a preparation method and application thereof. Background technique [0002] my country is a big country in the use of antibiotics, and the application of antibiotics is also very extensive. However, in recent years, driven by economic interests and lack of scientific knowledge, antibacterial drugs have evolved from widespread use to excessive use or even abuse, which seriously threatens human health. Rational use of antibacterial drugs and strengthening the supervision and management of antimicrobial drugs are gradually being concerned by the whole society. Penicillin is one of the most commonly used antibiotics in veterinary medicine. It is mainly used to prevent and treat mastitis in dairy cows, and to treat infections of the urinary tract, gastrointestinal ...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/48
Inventor 潘道东黄璐孙杨赢曾小群曹锦轩吴振李桦
Owner NINGBO UNIV
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