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Anti-streptomycin antibody and application thereof

A streptomycin and single-chain antibody technology, 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., to achieve Simple storage, high sensitivity, easy production effects

Inactive Publication Date: 2011-01-26
北京维德维康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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-streptomycin antibody and application thereof
  • Anti-streptomycin antibody and application thereof
  • Anti-streptomycin antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1, preparation and function test of antibody

[0058] 1. Preparation of streptomycin single chain antibody

[0059] (1) Antibody screening

[0060] 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 a complete set of primers, the heavy chain variable region (VH) and light chain variable region (VL) genes were respectively amplified using cDNA as a template. Splicing into a single-chain antibody gene (ScFv), then linking the ScFv with the vector pCANTAB5E to obtain pCANTAB5E / ScFv, and transforming Escherichia coli TGi to obtain a mouse-derived non-immune single-chain antibody library. Phage particles carrying multiple specific single-chain antibodies against streptomycin were screened by ELISA, and their corresponding nucleotide sequences were obtained by sequencing.

[0061] The single-chain antibody consists of a he...

Embodiment 2

[0090] Embodiment 2, the enzyme-linked immunosorbent assay kit that detects streptomycin and its preparation

[0091] 1. The enzyme-linked immunosorbent assay kit consists of the following materials:

[0092] 1. ELISA plate coated with streptomycin and carrier protein conjugate (STM-BSA);

[0093] 2. Streptomycin 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;

[0094] 3. Streptomycin standard product: streptomycin standard product is streptomycin sulfate, and the concentrations of the standard solution are 0μg / L, 0.5μg / L, 1.5μg / L, 4.5μg / L, 13.5μg / L, 40.5 μg / L; Streptomycin Sulfate was purchased from Sigma-Aldrich Company of the United States; the product catalog number is 46754; dilute to the above concentrations with sample diluent;

[0095] 4. Enzyme-labeled secondary antibody: h...

Embodiment 3

[0150] Embodiment 3, the test paper that detects streptomycin 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] The colloidal gold pad is coated with a colloidal gold-labeled streptomycin antibody (the protein shown in sequence 1 in the sequence listing);

[0155] Detection area and quality control area are arranged on described reaction film, and detection...

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Abstract

The invention discloses an anti-streptomycin antibody and application thereof. The antibody consists of a heavy chain variable region, a short peptide for connecting the heavy chain variable region with a light chain variable region, and a light chain variable region which are connected in turn, wherein an amino acid sequence of the heavy chain variable region is shown as No.1-No.123 amino acid residues from an N terminal in a sequence 1; and an amino acid sequence of the light chain variable region is shown as No.139-No.248 amino acid residues from the N terminal in the sequence 1. A kit anda colloidal 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, and can rapidly detect mass samples simultaneously and realize on-site high-throughput rapid detection. Therefore, the antibody, the kit, the colloidal gold test paper and the detection method have important significance for detecting streptomycin.

Description

technical field [0001] The present invention relates to a streptomycin antibody and its application. Background technique [0002] Streptomycin (STM) is one of the aminoglycoside antibiotics, which has significant antibacterial activity against Gram-negative bacteria and some Gram-positive bacteria, especially Mycobacterium tuberculosis. Streptomycin mainly acts on the ribosomes of cells and exhibits a bactericidal effect by inhibiting the biosynthesis of bacterial proteins. Because of its unique antibacterial activity, since streptomycin was first discovered in Streptococcus griseus in 1943, it has been widely used in the treatment of various diseases and animal feed additives, and has played an important role in the prevention and treatment of human and animal infectious diseases. had a huge effect. [0003] However, streptomycin has certain, sometimes even serious side effects on humans. The main side effect caused by streptomycin is ototoxicity. Because it can select...

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王奇昌师伟
Owner 北京维德维康生物技术有限公司
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