Anti-sulfonamides antibody and application thereof

A technology of sulfa drugs and single-chain antibodies, applied in the application, use of carriers to introduce foreign genetic material, cells modified by introducing foreign genetic material, etc., can solve the problems of complex production process, long time consumption, difficult operation, etc., and achieve storage Simple, sensitive, and easy-to-produce effects

Active Publication Date: 2011-01-26
北京英太格瑞检测技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation of monoclonal antibodies or polyclonal antibodies must be obtained through cell culture. The entire production process is complicated, time-consuming, expensive, and difficult to operate

Method used

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  • Anti-sulfonamides antibody and application thereof
  • Anti-sulfonamides antibody and application thereof
  • Anti-sulfonamides antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, preparation and function test of antibody

[0056] 1. Preparation of sulfonamide single-chain antibody

[0057] (1) Antibody screening

[0058] Six-month-old male Balb / c mice were taken, and the total RNA of splenocytes was extracted by Trizol one-step method, purified to obtain mRNA, and then reverse-transcribed to obtain cDNA. Using Oligo (dT) primers, cDNA was used as a template to amplify the heavy chain variable region (VH) and light chain variable region (VL) genes respectively, and the overlapping extension polymerase chain reaction (SOE-PCR) was used to pass a section The flexible peptide ((Gly4Ser)3-Linker) assembles VH and VL into scFv; the enzyme-cut scFv is linked into the ribosome display vector (pRDV), and the T7 promoter and ribosomes are introduced at the 5' end of the scFv by PCR The binding site, the spacer tolA sequence was introduced at the 3′ end to construct a ribosome display template, and the mRNA-ribosome-antibody ternary complex...

Embodiment 2

[0089] Embodiment 2, the enzyme-linked immunosorbent assay kit that detects sulfa drug and preparation thereof

[0090] 1. The enzyme-linked immunosorbent assay kit consists of the following materials:

[0091] 1. ELISA plates coated with sulfamethazine and carrier protein conjugates;

[0092] 2. Sulfa drug antibody: the single-chain antibody described in Example 1. The concentration of the antibody working solution is 5.0 ng / mL, and the antibody working solution is obtained by diluting the purified antibody in Example 1 with the sample diluent;

[0093] 3. Standard product of sulfa drug: the standard product of sulfa drug is sulfamethazine (SM 2 ), the concentrations of the standard solution were 0 μg / L, 0.5 μg / L, 1.5 μg / L, 4.5 μg / L, 13.5 μg / L and 40.5 μg / L; Sulfamethazine was purchased from Sigma-Aldrich, USA; The product catalog number is S6256; dilute to the above concentrations with sample diluent;

[0094] 4. Enzyme-labeled secondary antibody: horseradish peroxidase ...

Embodiment 3

[0150] Embodiment 3, the test paper that detects sulfa drug and its preparation and application

[0151] 1. The structure of the test paper

[0152] The test paper is composed of a sample absorbent pad, a colloidal gold pad, a reaction film and a water absorbent pad;

[0153] The sample absorption pad, the colloidal gold pad, the reaction membrane and the water absorption pad are sequentially connected, the end of the sample absorption pad is connected to the beginning of the colloidal gold pad, the end of the colloid gold pad is connected to the beginning of the reaction membrane, and the end of the reaction membrane is connected to the beginning of the reaction membrane. The beginning of the absorbent pad is connected;

[0154] Colloidal gold-labeled sulfa drug antibody is coated on the colloidal gold pad;

[0155] Detection area and quality control area are arranged on described reaction film, and detection area (C line) and quality control area (T line) all are the strip...

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Abstract

The invention discloses an anti-sulfonamides antibody and application thereof. The single-chain antibody consists of a heavy chain variable region, a short peptide for connecting the heavy chain variable region and a light chain variable region, and the light chain variable region connected in turn; the amino acid sequence of the heavy chain variable region is shown as the first position to the 118th position amino acid residues from the N terminal in a sequence 1; and the amino acid sequence of the light chain variable region is shown as the 134th position to 248th position amino acid residues from the N terminal in the sequence 1. A kit and a colloid gold test paper card have the advantages of high sensitivity, high accuracy, high precision, low cost, simple operation, short detection time, suitability for various units, simple storage and long quality guarantee period, can quickly detect massive samples simultaneously and can realize on-site high-flux quick detection. Therefore, the antibody, the kit, the colloid gold test paper and the detection method have important effect of detecting sulfonamides.

Description

technical field [0001] The invention relates to a sulfa drug antibody and application thereof. Background technique [0002] Sulfonamides are artificially synthesized sulfonamide derivatives, mainly used for the prevention and treatment of bacterial infectious diseases. Representative drugs are: sulfamethazine, sulfadiazine, sulfamethoxazole, sulfamethoxine, sulfaquinoxaline, etc. Sulfonamide drugs can inhibit Gram-positive bacteria and some negative bacteria, and can treat a variety of bacterial infections. They are widely used in veterinary clinics to treat various livestock and poultry diseases infected by sensitive bacteria. [0003] Due to the long time of action and metabolism of sulfa drugs in the body, sulfa drugs ingested through any route may accumulate in the human body. When the accumulated concentration exceeds a certain value, it will cause damage to the human body. Short-term high-dose or long-term low-dose stimulation can cause acute or chronic poisoning r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44C12N15/13C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10G01N33/566G01N33/577G01N33/531
Inventor 沈建忠江海洋吴小平王战辉李杰超徐飞刘丹王照鹏王进王世恩李艳伟李娜温凯
Owner 北京英太格瑞检测技术有限公司
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