Rabbit-derived anti-porcine parvovirus type 6 VP2 protein antibody and preparation method thereof

A parvovirus and protein technology, which is applied in the preparation methods of antiviral immunoglobulins, peptides, viruses/phages, etc., can solve the problems of low protein expression and inconvenient cultivation.

Pending Publication Date: 2021-07-23
开江县动物疫病预防控制中心
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, some scholars use eukaryotic expression system, baculovirus system, etc. to induce expression of VP2 protein, but the protein expression level is low, and it is not easy to cultivate.

Method used

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  • Rabbit-derived anti-porcine parvovirus type 6 VP2 protein antibody and preparation method thereof
  • Rabbit-derived anti-porcine parvovirus type 6 VP2 protein antibody and preparation method thereof
  • Rabbit-derived anti-porcine parvovirus type 6 VP2 protein antibody and preparation method thereof

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Embodiment Construction

[0034] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0035] 1. Materials and methods

[0036] 1. Materials

[0037] American type-porcine reproductive and respiratory syndrome virus (NA-Porcine Reproductive and Respiratory Syndrome Virus, NA-PRRSV) positive serum, porcine circovirus type 2 (Porcine circovirus type 2, PCV2) positive serum was purchased from VMRD Company; swine fever virus (Classical swine fever virus, CSFV) positive sera, Japanese encephalitis virus (Japanese encephalitis virus, JEV) positive sera, porcine pseudorabies virus (Porcine pseudorabies virus, PRV) positive sera were all provided by Kaijiang County Animal Disease Prevention and Control Center Prepare and save.

[0038] The clinical samples were 50 aborted sows, collected from different pig farms in 4 provinces in Southwest...

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Abstract

The invention discloses a rabbit-derived anti-porcine parvovirus type 6 VP2 protein antibody and a preparation method thereof. The preparation method comprises the following steps: designing a pair of specific primers according to a PPV6 epidemic strain genome sequence published by GenBank, constructing a pET30a-PPV6-VP2 recombinant expression vector, carrying out IPTG induced expression, Ni-NTA resin affinity chromatography purification and urea dialysis renaturation at different gradients, and performing uniform mixing and emulsification with a Freund's adjuvant in a ratio of 1: 1; and preparing PPV6 recombinant VP2 protein immunogen. A New Zealand white rabbit is subjected to multi-point subcutaneous injection on the back, hyperimmune serum is prepared, and the antibody is purified by adopting a saturated (NH4)2SO4 salting-out method. The prepared rabbit-derived anti-porcine parvovirus type 6 VP2 protein antibody is has good purity without bacteria, mycoplasma or exogenous virus pollution, and has good specificity without other common swine pathogen antibodies except the porcine parvovirus type 6 VP2 protein antibody, and the characteristics sufficiently meet the requirement for positive serum for exogenous virus detection, so that an important significance on healthy development of the veterinary biological product industry is achieved.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, and in particular relates to a preparation method of a rabbit-derived anti-porcine parvovirus type 6 VP2 protein antibody. Background technique [0002] Porcine parvovirus (PPV) belongs to the genus Parvovirus of the family Parvoviridae. It is an autonomously replicating virus and one of the main pathogens that cause reproductive disorders in sows. Sows infected with the virus can cause abortion and mummification. It caused serious economic losses to domestic and foreign pig industry; Mary and Mahncl first discovered the virus in 1966, and then developed to many countries and regions in the world; in the 1980s, many of our country Porcine parvovirus disease was isolated from all places. At present, with the development of molecular biology diagnostic techniques, several new porcine parvoviruses, such as PPV2, PPV3, PPV4, PPV5 and PPV6, have been discovered. Big difference. In 2014...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08C07K16/06C07K1/36C07K1/34C07K1/30C07K1/14C12N15/70C12N15/35
CPCC07K16/081C07K16/06C07K16/065C12N15/70C07K14/005C12N2750/14322C12N2750/14351
Inventor 欧云文潘琴张杰王勤刘俐君代军飞石义成任绍科邓书明
Owner 开江县动物疫病预防控制中心
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