Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

The anti-platelet membrane glycoprotein VI monoclonal antibody

A technology of membrane glycoproteins and platelets, applied in the direction of antibodies, anti-animal/human immunoglobulins, immunoglobulins, etc., can solve problems such as difficulty in obtaining high-titer human antibodies

Inactive Publication Date: 2006-10-25
MOCHIDA PHARM CO LTD
View PDF4 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There have been many reports on the preparation of human antibodies, but there are various problems in these reported methods. The current status is that a routine method suitable for all antigens has not been recognized, and it is usually difficult to obtain high-titer human antibodies

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
  • The anti-platelet membrane glycoprotein VI monoclonal antibody
  • The anti-platelet membrane glycoprotein VI monoclonal antibody
  • The anti-platelet membrane glycoprotein VI monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 7

[0055] Shown in embodiment 7.

[0056] The antibody of the present invention has the effect of inhibiting human platelet aggregation caused by collagen. Here, the platelet aggregation can be measured by a known method. For example, a platelet aggregation energy measuring device can be used, and the light transmittance is used as an index to calculate the aggregation rate. Usually, the aggregation rate at the point with the maximum light transmittance (hereinafter referred to as called the maximum aggregation rate). In the method described in Example 6 described later, the antibody of the present invention reduces the maximum aggregation rate at a concentration of preferably 10 μg / mL or less, more preferably 1 μg / mL or less, further preferably 0.1 μg / mL or less To preferably 50% or less of the control, more preferably 30% or less of the control, further preferably 20% or less of the control, most preferably 10% or less of the control. The method for measuring inhibition of co...

Embodiment 1

[0127] Example 1 Preparation of Human Soluble GPVI-Fc

[0128] A fusion protein (GPVI-Fc) of the extracellular domain of human GPVI and the Fc fragment of human IgG (GPVI-Fc) was prepared as an antigen for in vitro immunization and screening. The DNA manipulation was not particularly limited, and was carried out according to the second edition of Molecular Cloning (Molecular Cloning A Laboratory Manual 3rd., Joseph S., et al., Cold Spring Harbor Laboratory Press (2001)).

[0129] The GPVI-Fc expression plasmid was prepared by genetic engineering in the following order. First, using the plasmid pBK-CMV-GPVI-1 (Biochem Biophys. Res. Commun. 2000 August No. 14; 277(1): 27-36) incorporating the human GPVI cDNA cloned by Jiang Jiao et al. Sense primer-1 (SEQ ID NO.33, 5' end side contains restriction enzyme Xba I recognition sequence) and antisense primer-1 (SEQ ID NO.34, 5' end side contains restriction enzyme BamH I recognition sequence) PCR was performed to obtain cDNA encodin...

Embodiment 2

[0132] Example 2 Preparation of anti-GPVI monoclonal human antibody using human peripheral blood lymphocytes with anti-GPVI autoantibodies

[0133] The anti-GPVI monoclonal human antibody was prepared by fusing lymphocytes from blood donors confirmed to have GPVI autoantibodies with myeloma cells as follows. First, 6 ml of heparin-added blood obtained from aseptic blood collection from donors with written consent was placed in a Leucosep centrifugal separation tube (Greiner) added with 3 ml of Ficoll-Plus (Amersham Pharmacia Biotech AB), centrifuged at 1000 g, and lymphocytes were recovered components. The resulting lymphocyte fraction was washed twice with 800g centrifugation in Dulbecco's PBS (hereinafter referred to as D-PBS), then suspended in Hybridoma-SFM (Invitrogen) containing 10% FCS (fetal calf serum) to obtain 7.4 × 10 7 cells.

[0134] Add 2.5 μg / ml PHA-L (Sigma), 20 μg / ml LPS (DIFCO), and 10 μg / ml purified GPVI-Fc described in Example 1 to the above-mentioned ly...

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 present invention provides a human antibody or an active-fragment thereof that specifically binds to human platelet membrane glycoprotein VI and does not induce a human platelet aggregation independently; a cell that produces the antibody or its active-fragment; a pharmaceutical composition that comprises the antibody or its active-fragment as an active ingredient, and so on. The above-mentioned cell can be obtained for example, as follows: a peripheral-blood-lymphocyte of the human that produces an autologous antibody to GPVI is activated by in vitro immunization under specific conditions; a hybridoma with mouse myeloma cell is prepared; and then the hybridoma that produces a monoclonal antibody, which has a binding capacity to GPVI and has an activity that suppresses collagen-mediated agglutinability of the human platelet is selected.

Description

technical field [0001] The present invention relates to an antibody of human platelet membrane glycoprotein VI (hereinafter referred to as GPVI) and a cell producing the antibody. Background technique [0002] Platelets play an extremely important role in blood coagulation and body defense, and various pathological correlations can be elucidated from their physiological functions. It is particularly noticeable in the function of platelets to form hemostatic plugs. For example, when vascular endothelial cells are damaged, collagen, the main matrix protein under the vascular endothelium, is exposed, and platelets adhere to it. Next, platelets are activated by a signal from collagen, and finally, platelets are aggregated via fibrinogen. Depending on the case, this becomes a cause of a disease such as a thromboembolic disease, and attracts attention as a target of treatment. [0003] Conventionally, antiplatelet drugs such as aspirin, ticlopidine, and GPIIb / IIIa antagonists ha...

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): C12N15/13A61K39/395C07K16/18C07K16/28C12N5/10C12P21/02C12P21/08
CPCC07K16/2803C07K2317/56C07K2317/565C07K2317/92C07K2319/30C07K2317/76A61P13/12A61P27/02A61P3/10A61P7/02A61P7/04A61P9/00A61P9/04A61P9/10
Inventor 高山博史白川嘉门山川彻川原哲史
Owner MOCHIDA PHARM CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
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