Unlock instant, AI-driven research and patent intelligence for your innovation.

Anti-human XCR1 antibodies

An antibody, CDR2 technology, applied in the direction of antibodies, anti-animal/human immunoglobulins, anti-inflammatory agents, etc.

Inactive Publication Date: 2014-04-30
EISIA R&D MANAGEMENT CO LTD
View PDF5 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Reported high expression of XCR1 in placenta, but low expression in pancreas and thymus (Non-Patent Document 1)

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 XCR1 antibodies
  • Anti-human XCR1 antibodies
  • Anti-human XCR1 antibodies

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0510] (1) Preparation of mouse anti-human XCR1 monoclonal antibody

[0511] To obtain monoclonal antibodies against human XCR1, XCR1 knockout mice were immunized with membrane fractions of B300.19 cells expressing human XCR1. The membrane fraction was prepared according to the following steps: First, using a nitrogen gas cell disruption vessel (Parr Instruments), the suspension was dissolved in Ho buffer (0.25M sucrose, 10mM Hepes (pH7.4), 1mM EGTA, 0.5mM MgCl 2 , 1x Complete mini EDTA-free (Roche Applied Sciences) B300.19 cells expressing human XCR1 were disrupted (800 psi for 30 min on ice) and centrifuged (2,000 g for 10 min). The supernatant was collected and centrifuged again (100,000 g, 30 min). The pellet was suspended in 50 mM Hepes (pH 7.4) buffer and designated as the membrane fraction.

[0512] 160 μg or 260 μg of this membrane fraction was mixed with an equal volume of GERBU adjuvant (GERBU Biotechnik GmbH) and injected subcutaneously into the footpads of XCR1...

example 2

[0564] Preparation of chimeric anti-human XCR1 antibody and humanized anti-human XCR1 antibody

[0565] 5G7, which showed the highest neutralizing activity among 2H6, 5G7, and 11H2, was used to produce chimeric and humanized antibodies.

[0566] By combining (by overlap extension PCR) the gene sequence of the variable region of the 5G7 heavy chain and the constant region of human IgG2 (in which the V234A / G237A mutation was inserted into the heavy chain) and the sequence of the variable region of the 5G7 light chain and the constant region of human Igκ gene sequence, and the chimeric antibody was prepared by inserting the obtained sequence into an expression vector (pEE6.4 or pEE12.4). Tables 5 and 6 show the amino acid and nucleotide sequences of specific chimeric antibodies, respectively.

[0567] table 5

[0568] Amino acid sequence of chimeric anti-human XCR1 antibody

[0569]

[0570]

[0571] Table 6

[0572] Nucleic acid sequence of chimeric anti-human XC...

example 3

[0651] Pharmacological effects of mouse anti-XCR1 antibody

[0652] A mouse model of delayed contact dermatitis (DTH) was used to demonstrate the pharmacological effects of the anti-human XCR1 mouse monoclonal antibody (5G7) prepared in Example 2 above.

[0653] (1) Effect of mouse anti-human XCR1 antibody on ear swelling in DNFB (dinitrofluorobenzene)-sensitized mice

[0654] experimental method

[0655] 1. Sample Mice

[0656] Human XCR1 knock-in mice (mice whose XCR1 gene has been replaced by the human XCR1 gene) aged between 7 and 12 weeks on a C57BL / 6 background were used for the experiments.

[0657] 2. DNFB for sensitization and DNFB for induction

[0658] DNFB for sensitization and induction was prepared by mixing DNFB with a 4:1 mixture of acetone and olive oil to achieve a concentration of 0.5%. In addition, a 4:1 mixture of acetone and olive oil was used as a control solution for induction.

[0659] 3. DNFB administration method

[0660] The abdominal hair o...

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

An object of the present invention is to provide a monoclonal antibody binding to human XCR1, wherein the antibody binds to linear or discontinuous epitopes which comprise at least three amino acids selected from the group consisting of the 8th, 11th, 12th, 13th, 14th, 16th, 17th, 22nd, 23rd, 176th, and 177th amino acids in the amino acid sequence of SEQ ID NO: 91.

Description

technical field [0001] The present invention relates to antibodies that bind to human XCR1. Background technique [0002] Chemokines are a collective term for basic heparin binding proteins that have an effect on leukocyte chemotaxis and leukocyte activation. Based on a comparison of the primary structures of various chemokines, chemokines are classified into CXC, CC, C, and CX3C subfamilies according to the position of the conserved cysteine ​​residues. XCL1 (also known as lymphotactin (Ltn) or lymphotactin alpha (Ltn-α)) and XCL2 (also known as lymphotactin beta (Ltn-β)) are classified as Chemokines of subfamily C as described above. XCR1 (also known as GPR5, SCM-1α, or ATAC) is a G protein-coupled chemokine receptor that specifically binds to XCL1 and CXL2. [0003] Expression of XCR1 in various human tissues has been examined at the mRNA level. High expression of XCR1 in placenta was reported, but expression in pancreas and thymus was low (Non-Patent Document 1). Fu...

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): C07K16/28A61P37/00
CPCC07K2317/24C07K2317/55C07K16/2866C07K2317/34C07K2319/55C07K2317/76A61K2039/505A61P17/00A61P17/04A61P17/06A61P17/14A61P19/00A61P19/02A61P21/00A61P25/00A61P29/00A61P37/00A61P37/02A61P5/14A61P3/10C07K16/28
Inventor 坂本佳正西村美由希河野铁泽幸久今井俊夫
Owner EISIA R&D MANAGEMENT CO LTD