Monoclonal antibody for detecting aminoglycoside antibiotics and kit thereof

A monoclonal antibody and aminoglycoside technology, applied in the field of genetic engineering, can solve the problems of honey that cannot be exported to EU countries, loss of honey exporters and manufacturers, and side effects.

A monoclonal antibody and aminoglycoside technology, applied in the field of genetic engineering, can solve the problems of honey that cannot be exported to EU countries, loss of honey exporters and manufacturers, and side effects.

CN102776151AInactive Publication Date: 2012-11-14TARIM UNIV
2 Cites 5 Cited by

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Monoclonal antibody for detecting aminoglycoside antibiotics and kit thereof
  • Monoclonal antibody for detecting aminoglycoside antibiotics and kit thereof
  • Monoclonal antibody for detecting aminoglycoside antibiotics and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0023] Utilize the monoclonal antibody secreted by the monoclonal hybridoma cell line Pami-SP20 whose deposit number is CCTCC C201135 to obtain the detection method and kit for aminoglycoside antibiotics, and the experimental steps are as follows;

[0024] 1) Preparation of antibody detection plate:

[0025] Step 1, Carbodiimide (EDC) method, place EDC at room temperature for 3 hours; dissolve 10 mg BSA in 1 mL MES buffer, take 20 mg SM and dissolve in 2.5 mL MES buffer, and mix the two solutions ; Dissolve 5mg EDC in 0.5mL sterilized distilled water, slowly add the mixed solution of SM and BSA, react at room temperature for 6h, in a 4 ℃ refrigerator, use PBS 0.01 moL / L, pH 7.2, dialyze 4 times, each time For 5h, the coupling product was freeze-dried and stored for later use;

[0026] Step 2, immobilize the dialysis-purified complete antigen SM-BSA on an enzyme-linked plate, coat the enzyme-linked plate with a step concentration, 100ul per well, coat overnight at 4°C, and the...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a monoclonal antibody for detecting aminoglycoside antibiotics and a kit thereof. A hybridoma cell strain Pami-SP20 with the preservation number being CCTCCC201135 is acquired; andaccording to an aminoglycoside antibiotics residue quick-detection method established by a monoclonal antibody excreted by the strain, the monoclonal antibody excreted by the monoclonal hybridoma cell strain can be directly applied in aminoglycoside antibiotics remained ELISA (Enzyme-linked Immuno Sorbent Assay) quick-detection reagent preparation and detection of an acquired antibody remained by the aminoglycoside antibiotics through a double-antibody sandwich ELISA method. The aminoglycoside antibiotics detection kit comprises a detection plate wrapped by a streptomycin-BSA (Bovine Serum Albumin) antigen, the monoclonal antibody marked with HRP (Horse Radish Peroxidase), and ELISA reaction liquid. The monoclonal antibody excreted by the monoclonal hybridoma cell strainhas the advantages of strong specificity and high affinity, and provides a new manner for quick detection of aminoglycoside antibiotics remaining.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for preparing a hybridoma cell line of a general monoclonal antibody against aminoglycoside antibiotics, and obtaining a detection kit for its residue. Background technique [0002] Aminoglycoside antibiotics are composed of two or three amino sugar molecules and one amino cyclic alcohol connected by ether bonds, mainly including streptomycin, gentamicin, neomycin, tobramycin, kanamycin It is a broad-spectrum antibiotic with strong bactericidal effect on Mycobacterium tuberculosis, Leptospira, Actinomyces and various Gram-negative bacilli, and is used in animal husbandry production It is also often added to the feed to promote growth and development. At the same time, aminoglycoside antibiotics are prone to cross-resistance. For example, Escherichia coli, which was originally sensitive to streptomycin at a concentration of 1ug / ml, can tolerate a concentratio...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
14 Nov 2012
Publication
CN102776151A
IPC
C12N5/20; C07K16/44; G01N33/577; C12R1/91
Inventors
杨建军; 徐晓立