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Furazolidone metabolite derivative monoclonal antibody and applications thereof

A technology of monoclonal antibody and furazolidone, which is applied in the field of immunochemistry, can solve the problem that the actual residue of furazolidone cannot be determined, and achieve the effect of sensitive detection

Active Publication Date: 2013-02-20
深圳华利达植绒材料有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The actual residue of furazolidone cannot be determined only by detecting the parent drug

Method used

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  • Furazolidone metabolite derivative monoclonal antibody and applications thereof
  • Furazolidone metabolite derivative monoclonal antibody and applications thereof
  • Furazolidone metabolite derivative monoclonal antibody and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A. Reagent and material preparation

[0023] Furazolidone metabolite standard substance (AOZ, Shanghai Anpu Scientific Instrument Co., Ltd., NF005), furazolidone (AO, Institute for the Control of Pharmaceutical and Biological Products, 10046-200401); m-aldebenzoic acid (Alfa, B21277); DMF (sigma, D4551 ), sodium acetate (sigma, S2889), acetic anhydride (Sinopharm Chemical Reagent Co., Ltd.), bovine serum albumin (BSA) (Jackson, 001-000-173), ovalbumin (OVA) (Sigma, A5503), Freund's Complete Adjuvant (Sigma, F5881), Freund's Incomplete Adjuvant (Sigma, F5506), Tetramethylbenzidine (TMB) (Amresco, 0759), HAT (Sigma, H0262) and HT (Sigma, H0137) , sodium chloride (Amresco, 0241), potassium chloride (Amresco, 0395), potassium dihydrogen phosphate (Sigma, P9791), disodium hydrogen phosphate (Sigma, 71639), goat anti-mouse IgG-HRP (Jackson, 115- 035-044), dimethyl sulfoxide (DMSO) (Applichem, 0231), polyethylene glycol 4000 (PEG4000) (Sigma, P7306), DMEM high glucose medium ...

Embodiment 2

[0119] drug cross reaction test

[0120] According to the indirect competition ELISA method for monoclonal antibody screening established in Example 1, the monoclonal antibody of furazolidone metabolite derivatives and furazolidone hapten structural analogues such as nitrofurantoin, furaltadone, and furazolidone metabolite derivatives were tested for competition. These standard products were diluted into different concentrations for indirect competition ELISA, the inhibition curve was drawn, and the IC50 value and cross-reaction rate of the competitor were calculated. The results showed that the cross-reactivity of the monoclonal antibody with four structurally similar metabolic derivatives of furazolidone was very low, specifically The results are shown in Table 4.

[0121] type %CR type %CR Furazolidone metabolite derivatives 100% Nitrofurantoin metabolite derivatives <0.1%

Embodiment 3

[0123] This example is an example of the application of the monoclonal antibody 1C3' in the present invention in establishing an ELISA method for the detection of furazolidone metabolite residues, which can be used for the detection of furazolidone metabolite residues in muscle, aquatic products, and milk. Taking shrimp meat (purchased in the market) as an example, the main steps for detecting residues of furazolidone metabolites in shrimp meat are as follows:

[0124] 1. Sample preparation

[0125] (1) For shrimp meat sample, use a homogenizer to homogenize the sample, weigh 1.0±0.05 g of the homogenate (shrimp meat sample), and add it to a 15 mL centrifuge tube.

[0126] (2) Add 4.0 mL of distilled water, 0.5 mL of 1 M HCl and 100 μl of derivatization reagent, shake well.

[0127] (3) Incubate at 37°C for 16 h (overnight).

[0128] (4) Add 1.0 mL 0.5 M K 2 HPO 4 , 0.4 mL of 1 M NaOH, 2 g of NaCl, and 6 mL of ethyl acetate, and shake vigorously for 5 min.

[0129] (5) Ce...

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Abstract

The present invention relates to a monoclonal antibody and applications thereof, especially to a furazolidone metabolite derivative monoclonal antibody and applications thereof, and belongs to the technical field of immunochemistry. The furazolidone metabolite derivative monoclonal antibody 1C3' is produced by mouse hybridoma cell line 1C3, wherein a subtype is IgG1 type. According to the present invention, the monoclonal antibody 1C3' produced by the mouse hybridoma cell line 1C3 can be abundantly produced, and can be used to prepare an enzyme linked immunoassay kit and colloidal gold test paper strips for furazolidone metabolite (AOZ) detection so as to achieve a purpose of rapid and sensitive detection of furazolidone drug metabolites in muscle, liver and aquatic products and establish a foundation for furazolidone metabolite residue detection kit and colloidal gold product development.

Description

technical field [0001] The invention relates to a monoclonal antibody and its application, in particular to a monoclonal antibody of furazolidone metabolite derivatives and its application, and belongs to the technical field of immunochemistry. Background technique [0002] Furazolidone (FZ), also known as furazolidone, is a synthetic nitrofuran broad-spectrum antibacterial drug that can inhibit a variety of Gram-positive bacteria and Gram-negative bacteria, as well as some protozoa and fungi. Or killing effect, and stable efficacy, has been widely used in livestock and poultry and aquaculture. However, toxicological studies have found that furazolidone and its metabolite 3-amino-2-oxazolidinone (3-amino-2-oxazolidinone, AOZ) have strong toxic side effects, can cause cancer, mutagenesis, and can cause hemolytic Anemia, thrombocytopenic purpura, multiple neuritis and other diseases. In 1995, the European Union first prohibited the use of furazolidone in livestock and poultr...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44G01N33/577C12R1/91
Inventor 唐宏崔迎利王晓艳
Owner 深圳华利达植绒材料有限公司
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