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

Feline panleukopenia virus antibody sequence, tetrapeptide chain molecule, globulin molecule and application

A feline parvovirus and sequence technology, applied in the fields of feline parvovirus antibody sequence, immunoglobulin molecule and its application, and tetrapeptide chain molecule

Inactive Publication Date: 2020-11-10
青岛博隆基因工程有限公司
View PDF5 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The difficulty in solving the above problems and defects is as follows: in order to obtain an antibody library of a certain scale, the first thing to do is to use RT-PCR technology to obtain a complete set of antibody genes from the body. Verification of multiple pairs of primers for light and heavy chain variable region sequences

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
  • Feline panleukopenia virus antibody sequence, tetrapeptide chain molecule, globulin molecule and application
  • Feline panleukopenia virus antibody sequence, tetrapeptide chain molecule, globulin molecule and application
  • Feline panleukopenia virus antibody sequence, tetrapeptide chain molecule, globulin molecule and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Construction of cat-derived anti-feline parvovirus single-chain antibody library

[0063] 1. Isolation of Feline Peripheral Blood Lymphocytes

[0064] According to the International Animal Immunization Manual, 5 experimental cats were immunized with vaccine. Each immunization interval was 2 weeks, and a total of 3 immunizations were performed. Two weeks after the three immunizations, the peripheral blood of the cats was collected. Take 5-10mL of fresh blood and mix evenly with whole blood and tissue diluent at a ratio of 1:1-1:2. Add cell separation solution to a 15mL centrifuge tube, and gently add an equal volume of diluted anticoagulant blood along the tube wall. The horizontal centrifuge is centrifuged at a speed of 400g-800g and a time of 15min-25min. After centrifugation, the liquid in the centrifuge tube should be divided into four layers, from top to bottom: the first layer: plasma layer; the second layer: lymphocyte layer; The third layer: separat...

Embodiment 2

[0071] Example 2 Screening of cat-derived anti-feline parvovirus single-chain antibody library

[0072] 1. Preparation of Phage Antibody

[0073] Add 1 mL of phage primary antibody library to 3 mL of XLI-Blue bacterial solution with OD600=0.5, place at 37°C for 45 min, add 6 mL of LB liquid medium containing Amp+ (100 μg / mL), and add glucose (1mol / mL) at a ratio of 1:1000 L) Continue to cultivate, when the OD600 of the bacterial solution is 0.5, add the helper phage M13K07, place it in a 37°C incubator for 30min, and then incubate at 220rpm / min at 37°C for 1h, centrifuge the bacterial solution at 3500rpm for 10min, discard the supernatant, and use an equal volume LB liquid medium containing Amp+ (100 μg / mL) and Kana (50 μg / mL) was added with IPTG (1 mol / L) at a ratio of 1:1000, 220 rpm / min, 37°C, and shaken for 6 hours. Centrifuge the culture medium at 12000rpm for 10-20min, and collect the supernatant. Add PEG8000 at a ratio of 1:4-1:5, invert and mix well, at this time clo...

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

No PUM Login to View More

Abstract

The invention belongs to the technical field of virus antibodies, and discloses a feline panleukopenia virus antibody sequence, a tetrapeptide chain molecule, an immunoglobulin molecule and application, wherein the feline panleukopenia virus antibody sequence is a heavy chain variable region amino acid sequence SEQ ID NO: 1 and a nucleotide sequence SEQ ID NO: 3; and a light chain variable regionamino acid sequence SEQ ID NO: 2 and a nucleotide sequence SEQ ID NO: 4. A sequence screening method comprises the following steps of preparing a bacteriophage antibody; screening a bacteriophage antibody library; identifying an anti-feline panleukopenia virus feline-derived bacteriophage single-chain antibody by using a PhageELISA method; sending a scFv bacterial liquid with a positive PhageELISAidentification result to a sequencing company for sequencing to obtain variable region sequences of a heavy chain and a light chain of the feline-derived anti-feline panleukopenia virus genetic engineering antibody. The invention provides support for the construction of feline-derived anti-feline panleukopenia virus genetically engineered antibodies with high affinity and low immunogenicity. Themethod has important significance for promoting the development of feline-derived antibody drugs.

Description

technical field [0001] The invention belongs to the technical field of virus antibodies, and in particular relates to a feline parvovirus antibody sequence, a tetrapeptide chain molecule, an immunoglobulin molecule and applications thereof. Background technique [0002] At present, feline parvovirus (Feline panleukopenia virus) clinically manifests symptoms such as vomiting and diarrhea, enteritis, high fever, etc., can cause a sharp drop in the number of white blood cells, and can be transmitted through the respiratory tract, digestive tract, or direct contact. At present, prevention is still the main focus on feline parvovirus, and there is no specific treatment drug, and symptomatic treatment and supportive therapy are mostly used. [0003] At present, the clinical treatment of feline parvovirus disease mainly relies on monoclonal antibodies. Most of the commercially available antibodies are immune serum of xenogeneic animals. Although they can protect young animals and a...

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): C07K16/08G01N33/569C40B50/06
CPCC07K16/081C07K16/005G01N33/56983C40B50/06C07K2317/622C07K2317/56G01N2333/015
Inventor 曲雪婷尹燕博
Owner 青岛博隆基因工程有限公司
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