Anti-avian leukosis virus p27 protein monoclonal antibody, gold-colloidal strip containing same and application

A technology of avian leukosis virus and monoclonal antibody, which is applied in the fields of monoclonal antibody, gold standard test strip, and monoclonal antibody against p27 protein of avian leukosis virus, can solve the problem that there are no effective control measures for avian leukosis, and achieve easy promotion, High efficiency, clear and easy-to-discern results

Active Publication Date: 2015-12-30
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Avian leukemia has n

Method used

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  • Anti-avian leukosis virus p27 protein monoclonal antibody, gold-colloidal strip containing same and application
  • Anti-avian leukosis virus p27 protein monoclonal antibody, gold-colloidal strip containing same and application
  • Anti-avian leukosis virus p27 protein monoclonal antibody, gold-colloidal strip containing same and application

Examples

Experimental program
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Embodiment 1

[0025] Example 1 Preparation of Anti-Avian Leukosis Virus p27 Protein Monoclonal Antibody and Analysis of Monoclonal Antibody Specificity

[0026] 1 Materials and methods

[0027] 1.1 Virus strains, cells and experimental animals

[0028] ALV-J (HB09JY03) was isolated and identified by Harbin Veterinary Research Institute. ALV-J (HPRS103, GenBank: Z46390), ALV-A (RAV-1, GenBank: M19113) and ALV-B (RAV-2, GenBank: M14902) were donated by Professor Venugopal Nair, Pirbright Institute, UK; DF-1, S / P20 cells were preserved by our laboratory; 6-week-old BALB / c female mice were purchased from the Experimental Animal Center of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences.

[0029] 1.2 Main reagents

[0030] Escherichia coli DH5α and BL21 competent, prokaryotic expression vector pET-30a were preserved by our laboratory; pMD-18T vector and IPTG were purchased from Bao Biological Engineering (Dalian) Co., Ltd.; PEG-4000, HPR-labeled goat anti-mouse ...

Embodiment 2

[0063] Embodiment 2 detects the preparation and properties of the gold standard test paper strip of avian leukosis virus

[0064] 1.1 Preparation of colloidal gold solution

[0065] Colloidal gold particles with a particle size of 25 nm were prepared by sodium citrate reduction method. Under stirring, 100mL of 0.01% chloroauric acid aqueous solution was heated to boiling, and 1.5mL of 1% sodium citrate aqueous solution was quickly added, and the color of the solution was observed to change from white to black and then to wine red. When the color of the solution turns completely wine red, continue to heat for 5 minutes, then stop heating, cool naturally and store in the dark at 2-8°C.

[0066] 1.2 Preparation of gold-labeled conjugates

[0067] Gold conjugate labeling: add 0.1MK dropwise to 10mL colloidal gold solution 2 CO 3 0.1 mL, stirred for 5 min, added 200 μL of mouse monoclonal antibody 3H11 (prepared in Example 1) against avian leukosis virus p27 antigen, mixed for ...

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Abstract

The invention discloses an anti-avian leukosis virus p27 protein monoclonal antibody, a gold-colloidal strip containing the same, and application. The anti-avian leukosis virus p27 protein monoclonal antibody is generated by secretion of a hybridoma cell strain with a preservation number of CGMCC NO. 11089 or a hybridoma cell strain with a preservation number of CGMCC NO. 11090. The monoclonal antibody is high in titer to p27 protein and good in specificity, and therefore can be used for preparing a kit and the gold-colloidal strip for detecting an avian leukosis virus. A preparation method of the monoclonal antibody provided by the invention is simple, and an antibody purification process is simple, and the efficiency is high, and the cost is low. As the gold-colloidal strip prepared by the anti-avian leukosis virus p27 protein monoclonal antibody provided by the invention is adopted to detect the anti-avian leukosis virus, the specificity is strong, and the operation is simple, and convenience, speediness and simplicity are realized, besides, a special instrument and equipment are not needed, and professional training is not needed, and a result is clear and easy to recognize; the operation is simple and convenient; the popularization is easy, and the antibody is more suitable for real-time detection.

Description

technical field [0001] The invention relates to a monoclonal antibody, in particular to an anti-fowl leukosis virus p27 protein monoclonal antibody, a gold standard test strip containing the monoclonal antibody and its application, belonging to the field of biotechnology. Background technique [0002] Avian leukemia is a general term for a variety of tumor diseases in poultry caused by avian leukemia virus (ALV). ALV belongs to the Retroviridae family, the genus Avian Type C Retrovirus. According to the host range of virus infection, interference spectrum and envelope antigen, ALV can be further divided into seven subgroups A to G. Subgroup J avian leukemia virus (ALV-J) was first isolated in 1988 in the UK. The emergence of ALV-J was first reported in China in 1999, and then ALV infection appeared in most parts of China. The large-scale outbreak of ALV has brought great economic losses to the poultry industry in my country. [0003] There is currently no effective contr...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N5/20G01N33/577G01N33/569C12R1/91
Inventor 高玉龙王笑梅唐宏王牟平王晓艳朱海波祁小乐王永强高宏雷
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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