Anti-aflatoxin general type monoclonal antibody hybridoma cell line and application thereof

A hybridoma cell line, aflatoxin technology, applied in antifungal/algae/lichen immunoglobulin, biochemical equipment and methods, instruments, etc. problem, to achieve the effect of good detection sensitivity and affinity

Active Publication Date: 2014-08-27
无锡迪腾敏生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows us to create a special type of protein called avidinosity factor 19 which specifically targets bacteria causing food poisonings such as Escherichia coli. It does this through its ability to bind to specific molecules found on these harmful organisms' surfaces like DNA.

Problems solved by technology

This patented technical problem addressed in this patents relates to finding an effective way to measure chemicals called Toxyanotoxins (TET), specifically BTX, from various sources like crops or grapefruits without causing damage to humans or animals due to their presence. Current instruments used for measuring TET require complex operations and may be dangerous because these compounds were found only after being exposed during processing steps. Immune assays provide precise measurements but suffer from poor specificity and accuracy issues compared to other techniques.

Method used

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  • Anti-aflatoxin general type monoclonal antibody hybridoma cell line and application thereof
  • Anti-aflatoxin general type monoclonal antibody hybridoma cell line and application thereof

Examples

Experimental program
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Embodiment 1

[0021] Example 1: Preparation of hybridoma cell line 1G6

[0022] 1. Synthesis of complete antigen

[0023] Dissolve 1 mg of AFB1 (or AFM1) in 2 mL of methanol-water-pyridine (4:1:1, V / V / V), add 4 mg of carboxymethylhydroxylamine hydrochloride, and reflux at 85°C for 3 h , placed at room temperature for 16-24 h. Rotary evaporation, remove the organic solvent, add 1mL DMF to dissolve, then add 0.45 mg EDC and 0.3 mg NHS, mix well in the dark, stir at room temperature for 4 h; take another 5 mg KLH dissolved in 1 mL, 0.1 M, pH 8.0 in PBS solution; slowly add the above-mentioned activated aflatoxin solution dropwise into the KLH solution, stir and react at room temperature overnight, dialyze at 4°C for three days, and store in separate packages at -20°C.

[0024] 2. Animal immunity

[0025] Healthy BALB / c mice aged 6-8 weeks were selected for immunization. Take aflatoxin complete antigen (1 mg / mL) and the same amount of QuickAntibody immune adjuvant TM After mixing evenly, i...

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Abstract

The invention provides an anti-aflatoxin general type monoclonal antibody hybridoma cell line and application thereof, belonging to the field of food safety immunodetection. The cell line 1G6 provided by the invention is collected in the China General Microbiological Culture Collection Center (CGMCC) with a collection number of CGMCC No.7210. After an aflatoxin complete antigen is uniformly mixed with equivalent Quick Antibody immunologic adjuvant TM, an obtained mixture is injected to immunize BALB/c mice through leg muscle. B1-KLH (fumonisin B1-Keyhole Limpet Hemocyanin) is adopted for the first time of immunization, M1-KLH (fumonisin M1-Keyhole Limpet Hemocyanin) is adopted for the second time of immunization, and an M1 complete antigen (without containing adjuvant) is adopted for the last time of impaction immunization. Splenocytes of immunized mice are fused with myeloma cells through a PEG (polyethylene glycol) method, and the hybridoma cell line 1G6 is obtained through indirect ELISA (Enzyme-Linked Immuno Sorbent Assay) and indirect competence ELISA screening and three times of subcloning. A monoclonal antibody secreted from the cell line has better affinity and detection sensitivity for aflatoxins B1, B2, G1, G2 and M1, and can be used for immunodetection of the total quantity of the aflatoxins in the food safety.

Description

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Claims

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Application Information

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Owner 无锡迪腾敏生物科技有限公司
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