Gene engineering antibody for kitasamycin residue detection, and preparation method and application thereof

A technology of genetically engineered antibodies and kitasamycin, which is applied in the directions of genetic engineering, botany equipment and methods, biochemical equipment and methods, etc., can solve the problems of unobtained patent documents, achieve easy production, high detection accuracy, reduce The effect of fees
CN103288960AActive Publication Date: 2013-09-11HUAZHONG AGRICULTURAL UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
HUAZHONG AGRICULTURAL UNIVERSITY
Publication Date
2013-09-11

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Abstract

The invention discloses a gene engineering antibody for kitasamycin residue detection, and a preparation method and application thereof. The preparation method comprises the following steps: A. taking hybridoma cell strain KA / 2A9 preserved in liquid nitrogen, extracting splenocyte total RNA (ribonucleic acid) to obtain mRNA (messenger ribonucleic acid), and carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid); B. amplifying to obtain heavy chain variable region and light chain variable region genes; by using heavy chain and light chain antibody segments as primers, connecting the antibody heavy chain with the light chain by using a polypeptide chain; C. connecting digestion scFv into E.coliTG1, adding M13K07, and culturing to obtain a bacteriophage antibody library of kitasamycin-specific antibodies; D. adding the bacteriophage into E.coliHB2151 in a logarithmic growth phase, and carrying out IPTG (isopropyl-beta-D-thiogalactopyranoside) induction to obtain a specific bacteriophage antibody; and E. sequencing the verified Escherichia coli expression strain to obtain the single chain antibody amino acid and nucleotide sequences. The method is easy to implement, simple to operate and convenient for mass production of gene engineering antibodies. The invention is widely used for detecting kitasamycin residues in animal edible tissues, and provides residue level information of kitasamycin in animal edible tissues.
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Description

technical field

[0001] The invention belongs to the technical field of veterinary drug residue analysis and genetic engineering antibodies, and specifically relates to a genetic engineering antibody capable of detecting kitasamycin residues, and also relates to a preparation method of a genetic engineering antibody for kitasarmycin residue detection, and also relates to the kitasarmycin residue detection method. Application in genetically engineered antibodies for detection of protein residues. Background technique

[0002] Kitamycin is a macrolide antibiotic widely used in pig and poultry production. Kitamycin can cause adverse reactions in humans, mainly including mild gastrointestinal reactions such as diarrhea, nausea, vomiting, allergies, liver toxicity, tinnitus, hearing impairment and cardiotoxicity. Kitamycin residues can cause bacterial resistance in the human body and directly affect the treatment of human diseases. Bacterial drug resistance experiments in clinic...

Claims

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