Furaltadone metabolite derivative monoclonal antibody and applications thereof

A technology of monoclonal antibody and furanotazone, applied in the field of immunochemistry, can solve the problems of high operator requirements, high price, difficult to promote in grassroots units, etc., and achieve the effect of low cross-reaction rate.

Active Publication Date: 2013-02-20
深圳海路生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the expensive instruments required by the instrument method, it takes a long time, and has high requirements for operators, it is difficult to promote in grassroots units, and it is not suitable for rapid screening of high-throughput samples

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Reagent and material preparation

[0024] Furaltadone hydrochloride (AM, Shanghai Anpu Scientific Instrument Co., Ltd., C13963000) furaltadone metabolite (AMOZ, Shanghai Anpu Scientific Instrument Co., Ltd., CDEO-NF003); m-aldehyde benzoic acid (Alfa B21277); DMF ( sigma, D4551), bovine serum albumin (BSA) (Jackson, 001-000-173), ovalbumin (OVA) (Sigma, A5503), complete Freund's adjuvant (Sigma, F5881), incomplete Freund's 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 (Gibco, 11995), fetal bovine serum (Gibco, C20270).

[0025] Experimental animals and cells: Balb / c mice (6-8 weeks...

Embodiment 2

[0122] drug cross reaction test

[0123] According to the indirect competition ELISA method for monoclonal antibody screening established in Example 1, the monoclonal antibody of furaltadone metabolite derivatives and furaltadone hapten structural analogues such as nitrofurantoin, nitrofurazone, and furazolidone were subjected to competition tests. These standard products Dilute to different concentrations for indirect competition ELISA, draw inhibition curves, and calculate IC of competitors 50 The results showed that the cross-reactivity between the antibody and four structurally similar metabolic derivatives of furaltadone was very low. The specific results are shown in Table 4.

[0124] type %CR type %CR furaltadone metabolite derivatives 100% Nitrofurantoin metabolite derivatives <0.1%

Embodiment 3

[0126] This example is an example of the application of the monoclonal antibody 4B10' in the present invention in establishing an ELISA method for the detection of furaltadone metabolite residues, which can be used for the detection of furaltadone metabolite residues in muscle, aquatic products, and milk. Taking shrimp meat (purchased in the market) as an example, the main steps for detecting furaltadone metabolite residues in shrimp meat are as follows:

[0127] 1. Sample pretreatment

[0128] (1) Shrimp meat sample, homogenize the sample with a homogenizer, weigh 1.0±0.05 g of the homogenate (shrimp meat sample), and add it to a 15 mL centrifuge tube.

[0129] (2) Add 4.0 mL of distilled water, 0.5 mL of 1 M HCl and 100 μL of derivatization reagent, shake well.

[0130] (3) Incubate at 37°C for 16 h (overnight).

[0131] (4) Add 1.0 mL 0.5 M K 2 HPO 4 , 0.4 mL 1M NaOH, 2 g NaCl and 6 mL ethyl acetate, shake vigorously for 5 minutes.

[0132] (5) Centrifuge at 4000 rpm f...

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Abstract

The present invention relates to a monoclonal antibody and applications thereof, especially to a furaltadone metabolite derivative monoclonal antibody and applications thereof, and belongs to the technical field of immunochemistry. The furaltadone metabolite derivative monoclonal antibody 4B10' is produced by mouse hybridoma cell line 4B10, wherein a subtype is IgG1 type. According to the present invention, the monoclonal antibody 4B10' produced by the mouse hybridoma cell line 4B10 can be abundantly produced, and can be used to prepare an enzyme linked immunoassay kit and colloidal gold test paper strips for furaltadone metabolite detection so as to achieve a purpose of rapid and sensitive detection of furaltadone metabolites in muscle, milk and aquatic products.

Description

technical field [0001] The present invention relates to a monoclonal antibody and its application, especially furaltadone metabolites [0002] The derivative monoclonal antibody and its application belong to the technical field of immunochemistry. Background technique [0003] Nitrofuran drugs are synthetic broad-spectrum antibiotics, which have very good antibacterial effects and pharmacokinetic characteristics, and have been widely used as growth-promoting additives for livestock, poultry, and artificially farmed aquatic products. However, through long-term laboratory research, it has been found that nitrofuran drugs and metabolites can cause cancer and gene mutations in experimental animals. Cancer, poultry, and artificially raised aquatic products can also cause cancer and gene mutations, so these drugs are prohibited from being used in the treatment of animal diseases and in feed. [0004] my country issued a ban on the use of nitrofuran antibiotics in 2002. In the "L...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/577
Inventor 唐宏崔迎利王晓艳
Owner 深圳海路生物技术有限公司
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