Hybridoma cell strain secreting porcine circovirus type 2 Cap protein monoclonal antibody

A hybridoma cell line, porcine circovirus technology, applied in antiviral immunoglobulin, analytical materials, biochemical equipment and methods, etc.

Inactive Publication Date: 2014-04-02
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] PCV2 antibody detection methods include ELISA, IPMA and IFA, etc., virus antigen detection methods include virus isolation and identification, IPMA, IFA and PCR, etc., and their sensitivity and specificity are relatively high. Good, but the operation is time-consuming and labor-intensive, and requires high technology and equipment, so it cannot be promoted and applied
Although double-antibody sandwich ELISA has been reported in other pathogen detection, it has not been reported in PCV2 detection due to the limitation of antibodies

Method used

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  • Hybridoma cell strain secreting porcine circovirus type 2 Cap protein monoclonal antibody
  • Hybridoma cell strain secreting porcine circovirus type 2 Cap protein monoclonal antibody
  • Hybridoma cell strain secreting porcine circovirus type 2 Cap protein monoclonal antibody

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Experimental program
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Embodiment Construction

[0020] 1. Preparation of hybridoma cells

[0021] (1) Mice immunization: using the purified PCV2 virus liquid as the immunogen, subcutaneously inject 6-8 week-old female Balb / C mice at multiple points at a dose of 100 μg / mouse, and immunize 3 times with an interval of 3 weeks. After 10 days, blood was collected from the tail vein of the mice to determine the serum antibody titer by indirect ELISA method, and the mice with high titer were selected and injected intraperitoneally with 50 μg of PCV2 purified virus liquid for booster immunization 3 days before cell fusion.

[0022] (2) Determination of serum titer of immunized mice: The prokaryotic expressed PCV2CapC protein was used as ELISA coating antigen to detect the serum antibody titer of immunized mice, and the serum of non-immunized mice was set as a negative control. Finally, the maximum serum dilution of P / N ≥ 2.1 was used as the serum titer of mice.

[0023] (3) Cell fusion: Spleen cells of immunized mice were aseptica...

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Abstract

The present invention discloses a hybridoma cell strain secreting porcine circovirus type 2 Cap protein monoclonal antibody, and an application of the hybridoma cell strain in preparation of porcine circovirus type 2 Cap protein monoclonal antibody, wherein the hybridoma cell strain has the preservation number of CGMCC No.8169, and is preserved in the China general microbiological culture collection center. According to the present invention, the hybridoma cells have stable antibody secreting ability, the secreted monoclonal antibody provides good reaction specificity for porcine circovirus type 2 Cap protein, and the research establishes good material basis for researches on PCV2 etiology and PCV2pathogenic mechanism.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and in particular relates to a hybridoma cell line secreting a monoclonal antibody of porcine circovirus type 2 Cap protein and application thereof. Background technique [0002] Porcine circovirus type 2 (PCV2) is one of the important pathogens causing economic losses in pig-raising countries in the world. Clinically caused diseases are collectively referred to as porcine circovirus-associated disease (PCVAD) or porcine circovirus disease (PCVD), and include weaned pig multisystemic wasting syndrome (PMWS), reproductive failure, enteritis, proliferative necrotizing pneumonia, and porcine Respiratory disease complex (PRDC), among others, has brought huge losses to the swine industry worldwide. PCV2 has no envelope, and the PCV2 genome contains two main open reading frames, ORF1 and ORF2. ORF2 encodes a viral structural protein, the Cap protein. Cap protein is the main immunogenic protein of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/08G01N33/68G01N33/577G01N33/569
Inventor 姜平翟淑燕白娟
Owner NANJING AGRICULTURAL UNIVERSITY
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