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

Anti-human cd69 antibody, and use thereof for medical purposes

a technology of human cd69 and antibody, applied in the field of antihuman cd69 antibody, can solve the problems of substantially impossible inability to evaluate the pharmacological effect of existing antibodies in vivo, and the method of efficiently evaluating in vitro the pharmacological effect of an antibody to human cd69 is not availabl

Inactive Publication Date: 2015-04-30
GENEFRONTIER CORP +1
View PDF0 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides an anti-human CD69 antibody that can be used to treat allergic and inflammatory diseases. Additionally, an animal model is provided to evaluate the effectiveness of the antibody in an in-patent text describes the development of an anti-human CD69 antibody that can be used to treat allergic and inflammatory diseases. This treatment has shown promise in animal models, and a clinical trial is currently underway to determine its effectiveness in humans.

Problems solved by technology

However, since the ligand for CD69 is unknown, a method for efficiently evaluating in vitro a pharmacological effect of an antibody to human CD69 is not available.
Moreover, since existing antibodies to human CD69 do not cross-react with non-human CD69, evaluation of a pharmacological effect of the existing antibodies in vivo is substantially impossible, and even whether or not these antibodies afford a useful pharmacological effect is not clear.

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
  • Anti-human cd69 antibody, and use thereof for medical purposes
  • Anti-human cd69 antibody, and use thereof for medical purposes
  • Anti-human cd69 antibody, and use thereof for medical purposes

Examples

Experimental program
Comparison scheme
Effect test

example 1

Production of Antigen and Antibody

(1) Production of Human CD69 Recombinant Protein

[0112]Since human CD69 forms a homodimer via cysteine 68, cDNA (NM—001781) encoding the extracellular region (amino acid sequences; 62-199) containing cysteine 68 was inserted into a vector for Escherichia coli periplasm expression. Competent cells of Escherichia coli TG1F(−) strain prepared in advance (Z-competent E. Coli Transformation Buffer Set: manufactured by ZYMO RESEARCH) were transformed with this expression vector, and cultured on a LB agar plate containing chloramphenicol (final concentration 34 μg / mL) at 37° C. overnight. This Escherichia coli cells were inoculated in a 2×YT medium, and cultured at 37° C. for 3-5 hr (OD600=0.5-0.8). IPTG (final concentration 0.1 mM) was added, and the mixture was cultured at 25° C. overnight. The cultured Escherichia coli cells were collected by centrifugation, and lysed with lysis buffer (200 mM borate, 160 mM NaCl, 2 mM EDTA, 1 mg / ml lysozyme, pH 8.0), an...

example 2

Effect of Anti-Human CD69 Antibody on Intraalveolar Mononuclear Cell Infiltration

[0119]Mice obtained by crossing BALB / c mouse or CD69 deficient (CD69KO) mouse back-crossed not less than 10 times onto BALB / c (Murata, K. et al. 2003. Int. Immunol. 15: 987-992) with D011.10 transgenic mouse were used. The spleen CD4T cells of these mice were purified by AutoMACS sorter (Miltenyi Biotec) to a purity of >98%. The isolated CD4T cells were cultured with stimulation with immobilized anti-TCR and anti-CD28 monoclonal antibodies under Th2 conditions (IL-2 10 u / ml, IL-4 100 U / ml). Two days after the start of the culture, and human CD69 gene was introduced by a retrovirus vector containing human CD69 (hCD69) gene. The method of introducing human CD69 gene followed the method described previously (Kimura, M. et al. 2001. Immunity 15: 275-287). Five days after the start of the culture, the cultured cells were recovered, and the expression of human CD69 was confirmed by flow cytometry (FIG. 2). 50...

example 3

Selection of High Affinity Anti-Human CD69 Antibody

[0124]Selection of an antibody having higher affinity for human CD69 was tried by introducing a mutation into the light chain CDR3 of the antibodies selected in Example 1. To be specific, the methods described in Prassler J, Steidl S, Urlinger S. Immunotherapy. 2009 July; 1(4):571-83. and Hillig R C, Urlinger S, Fanghanel J, Brocks B, Haenel C, Stark Y, Sulzle D, Svergun D I, Baesler S, Malawski G, Moosmayer D, Menrad A, Schirner M, Licha K. J Mol Biol. 2008 Mar. 14; 377(1):206-19 were employed. The antibody selection round was repeated twice, the base sequences of the light chain CDR3 of the obtained antibody clones were examined and antibody clones having a novel sequence were identified. They were expressed in Escherichia coli, and ELISA was performed for the antigen by using the lysate and the amount of the antibody in the lysate was simultaneously measured by sandwich ELISA. The relative specific binding activity of each clone ...

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
concentrationaaaaaaaaaa
concentrationaaaaaaaaaa
concentrationaaaaaaaaaa
Login to View More

Abstract

The present invention provides an antibody that specifically binds to human CD69, has an activity to suppress allergic inflammation, and has cross-reactivity with mouse CD69. In addition, the present invention provides an antibody having high binding affinity for human CD69 and an activity to suppress allergic inflammations. The antibody of the present invention can be a human antibody.

Description

TECHNICAL FIELD[0001]The present invention relates to an anti-human CD69 antibody, and pharmaceutical use thereof.BACKGROUND ART[0002]CD69 is a type II transmembrane protein belonging to the C-type lectin family. Since the expression of CD69 increases within a few hours after stimulation of T cells and B cells, it is widely used as an early activation marker molecule to be an index of lymphocyte activation (non-patent document 1). In addition, the expression is also observed in T cells under selection during differentiation in thymus (non-patent documents 2 and 3). While CD69 is assumed to have a function as a coreceptor to potentiate signal transduction from an antigen receptor, the detail is unknown. Its ligand has not been identified to date. It is constitutively expressed in platelet, and the expression is also observed in activated neutrophils, eosinophils and the like. Therefore, it is assumed to play a role in the expression of function in platelets and topical inflammation r...

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(United States)
IPC IPC(8): C07K16/28
CPCC07K16/2851C07K2317/92C07K2317/33C07K2317/565A61K2039/505A61P29/00A61P37/08C07K2317/21C07K2317/34C07K2317/55
Inventor KOJOH, KANEHISAMIYAKOSHI, AKIRAKATOH, SHIZUETSUIHIJI, KUMIKOHAYAMI, YUKINAKAMURA, MIKIKONAKAYAMA, TOSHINORIIWAMURA, CHIAKI
Owner GENEFRONTIER CORP
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