Monoclonal antibody of chicken infectious laryngotracheitis virus gJ protein and application thereof

A technology of laryngotracheitis virus and monoclonal antibody, which is applied in the field of cell engineering, can solve problems such as difficulty in entering the clinical application stage, high requirements for test conditions and operators, and high test costs

Inactive Publication Date: 2011-08-17
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Among them, nucleic acid probe technology, polymerase chain reaction (PCR) technology and derived PCR-RFLP technology are widely used, but these methods have higher requirements for test conditions

Method used

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  • Monoclonal antibody of chicken infectious laryngotracheitis virus gJ protein and application thereof
  • Monoclonal antibody of chicken infectious laryngotracheitis virus gJ protein and application thereof
  • Monoclonal antibody of chicken infectious laryngotracheitis virus gJ protein and application thereof

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Embodiment 1

[0020] The preparation of embodiment 1 monoclonal antibody

[0021] 1. Purification and detection of recombinant protein

[0022] The induced protein rgJ was harvested after 5 hours, centrifuged at 6,000 rpm for 10 minutes, and 10 mL of binding buffer was added. Resuspend the bacterial liquid, place the sample in an ice bath and ultrasonically lyse (30% power) lyse for 3 seconds with an interval of 15 seconds, and sonicate until the liquid becomes transparent. Then centrifuge at 11,000rpm for 20min to remove cell debris, and transfer the supernatant to a new tube. Take the supernatant and precipitate separately for SDS-PAGE to determine whether the expressed protein is a soluble protein. The recombinant protein was purified using Bio-RAD's electroelution purification system, and the specific purification steps were as follows:

[0023] (1). Put on gloves and soak the membrane cup in the elution buffer at 60°C for at least 1 hour;

[0024] (2). Plug the filter plate to the ...

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Abstract

The invention discloses a monoclonal antibody of chicken infectious laryngotracheitis virus gJ protein and application thereof. The monoclonal antibody is screened to obtain a hybridoma cell strain (3B1E6F6) which can stably secrete the anti-gJ protein monoclonal antibody, wherein the preservation number of the microbe is China general microbiological culture collection (CGMCC) No. 4337. The monoclonal antibody secreted by the hybridoma cell strain can be reacted with the ILTV WG (laryngotracheitis virus WG) virulent strain and can not be reacted with the other correlated pathogenies of the birds. The monoclonal antibody of the invention can be used for detecting the chicken infectious laryngotracheitis virus, and laying a foundation for building a fast, simple and exact diagnosis method and distinguishing the infectious laryngotracheitis wild toxicity infection from the recombinant vaccine immunity.

Description

technical field [0001] The invention relates to a monoclonal antibody, in particular to a monoclonal antibody against chicken infectious laryngotracheitis virus gJ protein and a hybridoma cell strain secreting the anti-gJ protein monoclonal antibody, belonging to the field of cell engineering. Background technique [0002] Infectious laryngotracheitis (ILT) is an acute viral respiratory infectious disease of chickens, which breaks out worldwide, and the incidence rate can reach 100% when it is severe, and the fatality rate can be as high as 70%. The disease is caused by Infectious Laryngotracheitis Virus (ILTV). It is characterized by dyspnea, panting, coughing up bloody mucus, edema of the throat and trachea mucosa, bleeding and erosion, etc. It can cause infection in chickens. It is one of the important diseases that endanger the poultry industry (Saif, 2005). Since May and Tittster discovered the disease in Los Angeles, USA in 1925, it has spread all over the world. The...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/18G01N33/577
Inventor 王云峰石星明赵妍王玫
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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