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

Antibody binding to chondroitin sulfate proteoglycan-5

A chondroitin sulfate, proteoglycan technology, applied in the direction of antibodies, immunoglobulins, anti-animal/human immunoglobulins, etc., can solve problems such as short half-life

Pending Publication Date: 2021-02-26
KYOWA HAKKO KIRIN CO LTD +1
View PDF35 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, the antibody has a short half-life in the blood due to the systemic expression of the antigen (NPL12)

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
  • Antibody binding to chondroitin sulfate proteoglycan-5
  • Antibody binding to chondroitin sulfate proteoglycan-5
  • Antibody binding to chondroitin sulfate proteoglycan-5

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0548] [Example 1] Obtaining of anti-CSPG5 antibody

[0549] (1) Obtaining antibodies using human antibody phage library

[0550] VH gene fragments and VL gene fragments were amplified by PCR from cDNA derived from human PBMCs. The VH gene fragment and VL gene fragment were respectively inserted into phagemid vector pCANTAB 5E (manufactured by Amersham Pharmacia Biotech, Inc.), and plasmids were obtained by transforming Escherichia coli TG1 (manufactured by Lucigen Corporation). The resulting plasmid was infected with M13KO7 helper phage (manufactured by Invitrogen, Inc.), whereby a human antibody M13 phage library of VH gene and VL gene was obtained.

[0551] In addition, a synthetic human antibody M13 phage library incorporating random mutations in CDR3 was generated in the same manner.

[0552] Using the human antibody M13 phage library, the anti-CSPG5 monoclonal antibody was obtained using the following phage display method. The human CSPG5-FLAG_Fc or mouse CS...

Embodiment 2

[0560] [Example 2] Production of antibody

[0561] (1) Construction of CSPG5 scFv-hG4PE (R409K) expression vector

[0562] An expression vector for producing scFv-Fc antibodies in which each anti-CSPG5 scFv antibody was linked to a human IgG4 antibody comprising amino acid residue substitutions according to EU numbering S228P, L235E, and R409K (sometimes hereinafter) was constructed. referred to simply as the "IgG4 variant") Fc region.

[0563] Using the phagemid vector pCANTAB_CSPG5115 as a template, the gene fragment of the scFv region was amplified by PCR. The gene fragment for the hinge-CH2-CH3 region was amplified by PCR using a synthetic gene for the heavy chain constant region as a template. The obtained gene fragment was inserted into a pCI vector (manufactured by Promega, Inc.), thereby generating a pCI_CSPG5115scFv-hG4PE(R409K) vector.

[0564] Antibody expression vectors into which gene fragments of the scFv regions of each of the various types of anti-...

Embodiment 3

[0578] [Example 3] Analysis of reactivity with CSPG5 expressing cells

[0579] The nucleotide sequence encoding human CSPG5 is represented by SEQ ID NO: 159, the amino acid sequence deduced from the nucleotide sequence is represented by SEQ ID NO: 160, the nucleotide sequence encoding mouse CSPG5 is represented by SEQ ID NO: 161 represents, the amino acid sequence deduced from the nucleotide sequence is represented by SEQ ID NO: 162, the nucleotide sequence encoding monkey CSPG5 is represented by SEQ ID NO: 163, and the amino acid sequence deduced from the nucleotide sequence is represented by SEQ ID NO: 162 ID NO: 164 said.

[0580] The full-length gene sequences of human CSPG5, mouse CSPG5, and monkey CSPG5 were synthesized, and the gene sequences were respectively inserted into the BamHI-NotI sites of the pEF6 / V5-His (manufactured by Thermo Fisher Scientific, Inc.) vector, The following plasmid vectors for membrane expression of the corresponding types of CSPG5 were ...

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
particle diameteraaaaaaaaaa
Login to View More

Abstract

The present invention relates to: an antibody or a fragment of said antibody that binds to chondroitin sulfate proteoglycan-5 (CSPG5); a hybridoma that produces the antibody or the antibody fragment;a nucleic acid that contains a base sequence coding for the antibody or the antibody fragment; a transformed cell that contains a vector containing said nucleic acid; a method for manufacturing the antibody or the antibody fragment; a composition containing the antibody or the antibody fragment; a method for detecting or measuring an antigen present in the brain by employing the antibody or the antibody fragment; a method for diagnosing or treating a brain disease; a method for enhancing the retention of the antibody in the brain; a method for increasing the antibody quantity in the brain; andso forth.

Description

technical field [0001] The present invention relates to, for example, an antibody or antibody fragment thereof that binds to chondroitin sulfate proteoglycan 5 (CSPG5), a hybridoma producing the antibody or antibody fragment thereof, a nucleotide sequence encoding the antibody or antibody fragment thereof Nucleic acid, transformant cell comprising vector containing said nucleic acid, method for producing said antibody or antibody fragment thereof, composition comprising said antibody or antibody fragment thereof, and method for detecting or measuring antigen present in brain , a method for diagnosing or treating a brain disease, a method for improving the property of antibody accumulation in the brain, a method for increasing the amount of an antibody in the brain, etc., each of which uses the antibody or an antibody fragment thereof . Background technique [0002] Since the FDA approved the mouse anti-CD3 antibody murozumab-CD3 (OKT3) as the first antibody drug in 1986, ma...

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): C12N15/13A61K39/395A61K47/56A61K47/60A61K47/68A61K50/00A61K51/10A61P25/00A61P29/00A61P35/00C07K16/28C12N1/15C12N1/19C12N1/21C12N5/10C12P21/08
CPCA61P25/00A61P29/00A61P35/00A61K2039/505C07K16/3053C07K2317/52C07K2317/522C07K2317/524C07K2317/526C07K2317/31C07K2317/73C07K2317/77C07K2317/622A61K47/60A61K47/68A61K51/10G01N33/6857G01N33/6893C07K2317/565C07K2319/33C12N5/16G01N33/5058
Inventor 高桥信明中野了辅前田彩夏伊东祐二
Owner KYOWA HAKKO KIRIN CO LTD
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