Thiacloprid and acetamiprid monoclonal antibody hybridoma cell strain GW and application thereof

Abstract

The invention discloses a thiacloprid and acetamiprid monoclonal antibody hybridoma cell strain GW and application thereof, and belongs to the field of food security immunodetection. Thiacloprid and acetamiprid complete antigen of the strain is uniformly mixed with an equal amount of QuickAntibody-Mouse 5W adjuvant, and is injected to BALB / c mice through leg muscle. The dosage is 100mu g / mouse for the first time of immunization, the dosage is 50[mu]g / mouse for multiple times of intensified immunization, the immunization is implemented at an interval of 21 days, and thiacloprid and acetamiprid complete antigen (25[mu]g / mouse, without adjuvant) is adopted for immunization impact for the last time. Splenocyte of high-potency low IC50 mice is taken and fused with mouse myeloma cells by using a PEG method, indirect competitive inhibition enzyme-linked immunosorbent assay is adopted for screening, and three times of subcloning is implemented, so as to obtain the hybridoma cell strain. A monoclonal antibody secreted from the cell strain has relatively good specificity and detection sensitivity (the IC50 values are 0.1ng / mL and 0.4ng / mL respectively) for thiacloprid and acetamiprid, detection on the residual amounts of thiacloprid and acetamiprid in water, fruits and vegetables and cereals can be achieved, conditions are provided for immunodetection on thiacloprid and acetamiprid residues in food can be provided, and practical use values can be made.

Description

technical field [0001] The invention relates to a thiacloprid and acetamiprid monoclonal antibody hybridoma cell line GW and application thereof, belonging to the field of food safety immunology detection. Background technique [0002] Both thiacloprid and acetamiprid belong to the class of neonicotinoid insecticides, which are the action bodies of nicotinic acetylcholine receptors, and have been widely used in agricultural production. Thiacloprid is a low-toxicity insecticide, and acetamiprid It is a poisonous insecticide. These two drugs mainly act on the post-junction membrane of insect nerves. By combining with the nicotinic acetylcholine receptors, they interfere with the normal conduction of the insect nervous system, causing blockage of nerve channels, resulting in a large accumulation of acetylcholine, which makes the insects extremely excited, body convulsions, Paralyzed and died. It has strong contact, stomach poisoning and systemic effects. It is mostly used in...

AI Technical Summary

Problems solved by technology

It has been reported that exposure to nicotinic insecticides may have effects on the development of the nervous system in mammals

The European Union has restricted the use of neonicotinoid pesticides on some crops for nearly two years after a review by the European Food Safety Authority (EFSA) found they were harmful to bees

[0003] Chromatography is the most widely used method for detection of thiacloprid and acetamiprid, including gas chromatography (GC), high performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS) , supercritical fluid chromatography (SFC), capillary electrophoresis (CE) and other instrumental methods, but these methods require expensive instruments, professional operators, and the sample pretreatment is complex, costly, and time-consuming, and cannot achieve rapid processing of a large number of samples. Therefore, it is of great significance to establish a fast and simple detection method for thiacloprid and acetamiprid

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