Semicarbazide derivative monoclonal antibody and applications thereof

A monoclonal antibody, semicarbazide technology, applied in the field of immunochemistry, can solve the problems of time-consuming, unsuitable for rapid sample screening of high-throughput, high operator requirements, etc., and achieve the effect of low reaction rate

Inactive Publication Date: 2013-02-20
泰州康正生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the instruments required for these instrumental methods are expensive, time-consuming, and have high requirements for oper

Method used

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  • Semicarbazide derivative monoclonal antibody and applications thereof
  • Semicarbazide derivative monoclonal antibody and applications thereof
  • Semicarbazide derivative monoclonal antibody and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Reagent and material preparation

[0023] Furacillin (Shanghai Anpu Scientific Instrument Co., Ltd., C15571000), SEM (Shanghai Anpu Scientific Instrument Co., Ltd., C16933500), p-aldehyde benzoic acid (Sigma 124 91-5), 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), tetramethyl benzidine (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), dimethylsulfoxide (DMSO) (Applichem, 0231), polyethylene glycol 4000 ( PEG4000) (Sigma, P7306), DMEM high glucose medium (Gibco, 11995), fetal bovine serum (Gibco, C20270).

[0024] Experimental animals and cells: Balb / c mice (6-8 weeks old, female) were purchased from the An...

Embodiment 2

[0124] drug cross reaction test

[0125] According to the indirect competition ELISA method for monoclonal antibody screening established in Example 1, the monoclonal antibody of semicarbazide and semicarbazide hapten structural analogs such as metabolite derivatives of nitrofurantoin, furaltadone, and furazolidone were carried out in a competition test, and these metabolic Dilute the standard substance derivatives into different concentrations for indirect competition ELISA, draw the inhibition curve, and calculate the IC of the competitor 50 The results showed that the cross-reaction rates between the antibody and the other three furan metabolite derivatives were less than 0.1%. The specific results are shown in Table 4.

[0126] Table 4 Cross-reaction rate between semicarbazide derivatives and other furan drug derivatives

[0127] type %CR type %CR Furacilin 100% Nitrofurantoin metabolite derivatives <0.1%

Embodiment 3

[0129] This example is an example of the application of the monoclonal antibody 9B10' in the establishment of an ELISA method for detecting residues of nitrofurazone metabolites in the present invention, which can be used to detect residues of nitrofurazone metabolites in honey and animal tissues. Taking shrimp meat (purchased in the market) as an example, the main steps to detect residues of nitrofurazone metabolites in shrimp meat are as follows:

[0130] 1. Sample pretreatment

[0131] (1) Homogenize the shrimp sample with a homogenizer, weigh 1.0±0.05 g of the homogenate, and add it to a 15 mL centrifuge tube;

[0132] (2) Add 4.0 mL of distilled water, 0.5 mL of 1 M HCl and 100 μL of derivatization reagent, shake well.

[0133] (3) Incubate at 37°C for 16 h (overnight);

[0134] (4) Add 1.0 mL 0.5 M K2HPO4, 0.4 mL 1 M NaOH, 2 g NaCl and 6 mL ethyl acetate respectively, shake vigorously for 5 min;

[0135] (5) Centrifuge at 4000 rpm for 10 min at room temperature (20-25...

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Abstract

The present invention relates to a monoclonal antibody and applications thereof, especially to a semicarbazide (nitrofural metabolite) derivative monoclonal antibody and applications thereof, and belongs to the technical field of immunochemistry. The semicarbazide derivative monoclonal antibody 9B10' is produced by mouse hybridoma cell line 9B10, wherein a subtype is IgG1 type. According to the present invention, the monoclonal antibody 9B10' produced by the mouse hybridoma cell line 9B10 can be abundantly produced, and can be used to prepare an enzyme linked immunoassay kit and colloidal gold test paper strips for semicarbazide residue detection so as to achieve a purpose of rapid and sensitive detection of semicarbazide residues in honey and animal tissues.

Description

technical field [0001] The invention relates to a monoclonal antibody and its application, in particular to a semicarbazide derivative monoclonal antibody and its application, and belongs to the technical field of immunochemistry. Background technique [0002] Nitrofurazone belongs to nitrofuran drugs and is a broad-spectrum antibacterial drug. Because of its low price and good effect, it is used as a therapeutic drug and feed additive to control diseases in livestock, poultry and aquaculture. The pharmacophore-nitro group in the nitrofurazone molecule is the main reason for its pharmacological action and toxicity, and it also makes nitrofurazone have side effects such as teratogenicity, carcinogenicity, and induction of chromosome damage in mammalian cells. The European Union listed nitrofurazone as a prohibited drug in 1995. In Announcement No. 193 issued by the Ministry of Agriculture of China in April 2002, nitrofuran was included in the "List of Veterinary Drugs and Ot...

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 泰州康正生物技术有限公司
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