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Clenbuterol hydrochloride micromolecule hapten high-sensitivity detection test paper based on nano antibody, and preparation method thereof

A clenbuterol hydrochloride, nanobody technology, applied in biological testing, chemical instruments and methods, anti-animal/human immunoglobulin, etc., can solve problems such as limitations

Active Publication Date: 2020-06-02
XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, LC-MS, GC-MS and ELISA methods require professional and technical personnel to operate and rely on various instruments and equipment, so they are usually limited to laboratory use

Method used

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  • Clenbuterol hydrochloride micromolecule hapten high-sensitivity detection test paper based on nano antibody, and preparation method thereof
  • Clenbuterol hydrochloride micromolecule hapten high-sensitivity detection test paper based on nano antibody, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0062] (2) Preparation of anti-clenbuterol hydrochloride nanobody

[0063] (2.1) Alpaca immunity and blood collection

[0064] At the first immunization, 400 μg of BSA-CL mixed with complete Freund's adjuvant was injected through the cervical lymph nodes of the alpaca. Two weeks later, BSA-CL was mixed and emulsified with Freund's incomplete adjuvant, and the second immunization was carried out as above, and a total of 4 immunizations were carried out. Blood was collected from the neck vein, the antibody was detected by Western blot, and the antibody titer was determined by indirect ELISA established with OVA-CL as the coating antigen. Peripheral blood lymphocytes were isolated, total RNA was extracted, mRNA was purified, and first-strand cDNA was synthesized by reverse transcription.

[0065] (2.2) Preparation of phage peptide library

[0066] The PCR reaction was carried out by using the primers that specifically amplified the variable regions of alpaca IgG2 and IgG3 heav...

Embodiment 1

[0108] For the detection of small molecules of clenbuterol hydrochloride in pig urine samples, the urine samples were collected in a clean container and can be detected directly. If there is turbidity, centrifuge at 3000r / min for 5min, and take the supernatant for testing. Use clenbuterol hydrochloride small-molecule hapten high-sensitivity detection test paper to detect and judge the results according to the above-mentioned operation method (13) to determine whether the pigs eat clenbuterol hydrochloride small-molecule drug. Two reddish-brown bands (test line and quality control line) appear on the test paper. "||" is negative for clenbuterol hydrochloride, which means that the pigs to be tested have not consumed the small-molecule drug clenbuterol hydrochloride; only one reddish-brown band appears The band (quality control line) "|" is positive for clenbuterol hydrochloride, indicating that the pigs to be tested have eaten the small molecule drug clenbuterol hydrochloride; t...

Embodiment 2

[0110] For the detection of small molecules of clenbuterol hydrochloride in raw pork samples, take about 5 grams of meat slurry samples and place them in a centrifuge tube, boil them for 10 minutes, and take the supernatant for detection. If there is turbidity, centrifuge at 3000r / min for 5min, and take the supernatant for testing. Use clenbuterol hydrochloride small-molecule hapten high-sensitivity detection test paper to detect and judge the results according to the above-mentioned operation method (13) to determine whether the pigs eat clenbuterol hydrochloride small-molecule drug. Two reddish-brown bands (test line and quality control line) "||" on the test paper are negative for clenbuterol hydrochloride, indicating that the pigs to be tested have not consumed the small-molecule drug clenbuterol hydrochloride; one reddish-brown band ( Quality control line) "|" is positive for clenbuterol hydrochloride, indicating that the pigs to be tested have eaten the small molecule dr...

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Abstract

The invention discloses clenbuterol hydrochloride micromolecule hapten high-sensitivity detection test paper based on a nano antibody, and a preparation method of the clenbuterol hydrochloride micromolecule hapten high-sensitivity detection test paper. According to the clenbuterol hydrochloride micromolecule hapten high-sensitivity detection test paper, a gold-labeled pad in the detection test paper adsorbs a colloidal gold nanoparticle labeled anti-clenbuterol hydrochloride small molecule hapten nano antibody Nb; a detection line T on a detection membrane is a conjugate BSA-CL blot of clenbuterol hydrochloride micromolecule CL and bovine serum albumin BSA, and a quality control line C on the detection membrane is an anti-BSA protein rabbit-derived polyclonal antibody pAb blot. The invention further specifically discloses a preparation method of the clenbuterol hydrochloride micromolecule hapten high-sensitivity detection test paper based on the nano antibody. The clenbuterol hydrochloride micromolecule hapten high-sensitivity detection test paper based on the nano antibody prepared by adopting the preparation method is simple and convenient in detection operation, high in specificity, high in sensitivity, capable of rapidly giving a visual result and easy to popularize and apply in life practice.

Description

technical field [0001] The invention belongs to the technical field of high-sensitivity detection equipment for determining food safety, and in particular relates to a nanobody-based clenbuterol hydrochloride small-molecule hapten high-sensitivity detection test paper and a preparation method thereof. Background technique [0002] Immunochromatography test paper detection technology is a new light and simplified detection technology developed based on monoclonal antibody technology, immune labeling technology and immune chromatography technology. It can be used for qualitative, semi-quantitative and quantitative detection of antigens, antibodies and haptens. Simple, specific, fast and sensitive. Compared with analytical methods such as high performance liquid chromatography and gas chromatography, test paper detection technology does not rely on professional technicians and expensive instruments and equipment, and is less costly and time-consuming. The types of samples test...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/532C07K16/18C07K16/06C07K16/44
CPCG01N33/94G01N33/558G01N33/532C07K16/18C07K16/06C07K16/44C07K2317/569C07K2317/14Y02A50/30
Inventor 王寅彪王向鹏陈礼朋王磊吴卫东夏小静郭军庆乔松林
Owner XINXIANG MEDICAL UNIV
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