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Preparation method of PCV2Cap protein monoclonal antibody

A monoclonal antibody and protein technology, applied in the direction of antiviral immunoglobulin, etc., can solve the problems of time-consuming, difficult cultivation, cumbersome preparation process of cap protein monoclonal antibody, etc., and achieve short preparation time, simple operation and high titer Effect

Inactive Publication Date: 2014-03-26
JIANGSU NANNONG HI TECH
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Problems solved by technology

At present, the cultivation of PCV2 is relatively difficult. The preparation process of cap protein monoclonal antibody is very cumbersome and time-consuming. The titer of the monoclonal antibody produced is low, which cannot meet the needs of the experiment.

Method used

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Effect test

Embodiment Construction

[0011] The preparation of the PCV2Cap protein monoclonal antibody of the present invention comprises the following steps:

[0012] Step 1. Animal immunization

[0013] The extracted Cap protein was diluted to 1 mg / mL, mixed with an equal volume of Freund's complete adjuvant, and fully emulsified, and 5 5-week-old Balb / c mice were subcutaneously inoculated at multiple points, with an antigen dose of 50 μg / mouse. After 2 weeks, the same dose of antigen treated with Freund's incomplete adjuvant was injected, and the antigen was injected every 2 weeks, repeated 3 times. 3-5 days before cell fusion, intraperitoneal injection of Cap protein without adjuvant for booster immunization, and then cell fusion can be carried out.

[0014] Step 2: Establishment of Cap protein monoclonal antibody screening method

[0015] A) Establishment of Cap protein indirect ELISA method

[0016] (1) Dilute the extracted Cap protein with carbonate buffer (pH 9.6) to 1, 5, 10, 20, 30 μg / mL, 100 μL / well...

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Abstract

The invention relates to a preparation method of a PCV2Cap protein monoclonal antibody. The preparation method comprises the steps of immunizing an animal, fusing cells, selecting positive hybridoma cells and colonizing. By adopting the preparation method, the preparation time is short, operation is simple, the valence of the extracted antibody is high, and the experimental requirement can be met.

Description

technical field [0001] The invention relates to the preparation of a PCV2Cap protein monoclonal antibody. Background technique [0002] Porcine circovirus (PCV) disease is a new and important infectious disease affecting the pig industry in the world, and has caused serious economic losses to the pig industry in my country. PCV has two serotypes, namely PCV2 1 and PCV2, among which PCV 1 is a non-lethal virus, while PCV2 is a lethal virus. PCV2 contains two main open reading frames, ORF1 and ORF2, which encode viral replication-associated protein (Rep) and viral capsid protein (Cap), respectively. Compared with PCV2, PCV1 and PCV2, the amino acid sequence homology of Rep protein reached 86%, which is also the main reason for the cross-reactivity of antigens between the two PCVs. Cap is the main structural protein of the virus, which contains the main antigenic epitopes of the virus, and is an important antigen to stimulate the body to generate an immune response. The amino...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08
Inventor 季伟缪芬芳毛晓娜
Owner JIANGSU NANNONG HI TECH
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