Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Purification method of virus-like particles of porcine parvovirus and application of purification method

A technology of parvovirus and purification method, which is applied to the preparation methods of peptides, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of low added value of products and increased production costs, and achieves the effect of being beneficial to production operations.

Active Publication Date: 2019-09-27
扬州优邦生物药品有限公司
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Especially for veterinary vaccines, the added value of the product itself is low. If the purification process with cumbersome steps is adopted, the production cost will inevitably increase greatly
[0004] At present, there is no technology for the industrial purification of porcine parvovirus virus-like particles

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Purification method of virus-like particles of porcine parvovirus and application of purification method
  • Purification method of virus-like particles of porcine parvovirus and application of purification method
  • Purification method of virus-like particles of porcine parvovirus and application of purification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The preparation process of embodiment 1 porcine parvovirus semi-finished product

[0023] The nucleotide sequence encoding the porcine parvovirus virus-like particle gene is connected to the baculovirus to construct a recombinant baculovirus seed, and the virus seed infects insect cells (Sf9 cells), and the culture conditions of Sf9 cells are 27°C, 140rpm, 96h~ 120h. When the cells were lysed to release the virus, the venom was collected and the porcine parvovirus virus-like particles were inactivated by binary ethylenimine (BEI).

Embodiment 2

[0024] Example 2 Purification of porcine parvovirus virus-like particles

[0025] The inactivated venom is clarified to remove the precipitate, the pH of the semi-finished product is adjusted to 5.5-6.0 with 2% acetic acid, and the semi-finished product before purification of porcine parvovirus virus-like particles is obtained.

[0026] 1) weak anion exchange chromatography

[0027] The pretreated semi-finished product was added to a weak anion exchange column (DEAE Focusose 6XL, Wuhan Huiyan Biotechnology Co., Ltd.) equilibrated with sodium acetate buffer A (20mM sodium acetate, 0.1M NaCl, pH=5.5), Use sodium acetate buffer B (20mM sodium acetate, 1.0M NaCl, pH=5.5) for elution, the elution volume is 3 column volumes, the flow rate is 2.0mL / min, and the peak components are collected, and the weak anion exchange Chromatograms such as figure 1 shown.

[0028] 2) Determination of active protein components by polyacrylamide gel electrophoresis (SDS-PAGE) Take 80 μL of samples ...

Embodiment 3

[0050] Example 3 Safety Test of Vaccines Containing Porcine Parvovirus Virus-like Particles

[0051] 1. The same amount of adjuvant was added to the samples before and after the purification of the experimental vaccine porcine parvovirus virus-like particles to prepare the vaccine. The vaccine batch before purification was defined as VP2-01, and the vaccine batch after purification was defined as VP2-02.

[0052] 2. Test piglets were purchased from Yunli Animal Husbandry Co., Ltd., Dantu District, Zhenjiang City, and all of them were negative for porcine parvovirus antigen antibodies.

[0053] 3. Experimental method

[0054] 3.1 Single-dose vaccination safety experiment

[0055] 3.1.1 Safety test of piglets

[0056] 20 weaned piglets aged 3 to 5 weeks, 5 piglets in each batch of vaccine, intramuscular injection in the neck, 2mL / head, and the remaining 5 piglets as a control, inoculated with normal saline, carefully observed the reaction of the pigs after injection, and obser...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
recovery rateaaaaaaaaaa
Login to View More

Abstract

The invention discloses a purification method of virus-like particles of porcine parvovirus and an application of the purification method, and belongs to the field of separation and purification technology of biological products. The method comprises the following steps: clarifying a culture of expressed virus-like particles of the porcine parvovirus, collecting supernatant, and performing inactivation with pyrrole to obtain a semi-finished product; regulating the pH value of the semi-finished product to be smaller than 7; adding the regulated semi-finished product to a weak anion exchanging column balanced by a first buffer solution, eluting the weak anion exchanging column with a second buffer solution, and collecting peak components; determining active protein components of each peak by polyacrylamide gel electrophoresis. The purity of the virus-like particles of the porcine parvovirus obtained with the purification method is up to 90%, and recovery rate is up to 99%. The method is conductive to production and operation, and a safer and more reliable way is provided for preparing inactivated vaccine for rhabdovirus vectors of the porcine parvovirus.

Description

technical field [0001] The invention relates to a purification method and application of porcine parvovirus virus-like particles, belonging to the technical field of separation and purification of biological products. Background technique [0002] Porcine parvovirus (PPV) belongs to the genus Parvovirus of the family Parvoviridae, and is an autonomously replicating virus. Clinically, it is characterized by a series of symptoms such as abortion, stillbirth, deformed fetus, fetal mummification and weak offspring in pregnant sows. . Porcine parvovirus is often co-infected with other pathogens that cause reproductive disorders in sows to cause disease, which has brought serious losses to the pig industry. [0003] So far, the vaccines commonly used in China are mainly inactivated whole virus vaccines and recombinant subunit vaccines expressed by baculovirus. Compared with inactivated whole virus vaccines, subunit vaccines have the advantage of high antigen expression, but Ther...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/015C07K1/18C12N15/866A61K39/125A61P31/20
CPCA61K39/12A61K2039/5252A61K2039/552A61P31/20C07K14/005C12N15/86C12N2710/14043C12N2750/14023C12N2750/14034C12N2750/14051
Inventor 范娟丁国伟李群潘杰徐萍叶正琴李甜甜魏荣荣孙舒李琛
Owner 扬州优邦生物药品有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com