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

Use of the endoglycosidase Endos for treating immunoglobulin g mediated diseases

A technology of use and disease, applied to the use field of endoglycosidase EndoS for the treatment of diseases mediated by immunoglobulin G, and can solve problems such as no discovery

Active Publication Date: 2013-08-28
ハンサバイオファルマアクチボラゲット
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No other substrates of EndoS have been found so far

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
  • Use of the endoglycosidase Endos for treating immunoglobulin g mediated diseases
  • Use of the endoglycosidase Endos for treating immunoglobulin g mediated diseases
  • Use of the endoglycosidase Endos for treating immunoglobulin g mediated diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] Example 1: EndoS efficiently hydrolyzes IgG in human blood

[0115] To be effective as a therapeutic against pathological IgG, EndoS needs to be active at low concentrations in the context of human whole blood. To investigate this, recombinant EndoS (rEndoS) without a signal sequence (ie, having the sequence of SEQ ID NO: 1 ) was produced and purified as previously described (Collin & Olsén, 2001, Infect. Immun. 69:7187-7189). Increasing final concentrations (0, 0.31, 0.63, 1.25, 2.5, 5, 10 and 20 μg / ml) of rEndoS were rolled end over end in 500 μl of heparinized human blood from healthy volunteers at 37°C Incubate with rotation for 1 hour. Samples were centrifuged at 720 xg for 10 minutes at 4°C, after which IgG in plasma was purified using protein G sepharose according to the manufacturer's instructions (GE Healthcare Biosciences, Uppsala, Sweden). Consistent with previous findings on the IgG binding proteins protein H (from S. pyogenes) and protein A (from Staphy...

Embodiment 2

[0118] Example 2: EndoS efficiently hydrolyzes IgG in rabbits

[0119] To further substantiate the utility of EndoS as a therapeutic agent, the IgG glycan hydrolytic activity of EndoS in the circulation of living animals was studied. Swedish loop rabbits weighing approximately 3 kg were injected intravenously with 1 mg of rEndoS, corresponding to an rEndoS:IgG ratio of approximately 1:2000 (assuming that rEndoS is only distributed in the blood). Animals showed no signs of disease. Serum samples were collected at 0, 1, 2, 4, 6, 8 and 12 hours and on days 1, 2, 3, 4, 5, 6, 8 and 10. Serum IgG was analyzed for glycosylation status by SDS-PAGE and lectin blot analysis as described previously for human blood.

[0120] These experiments showed that, prior to injection, the apparent molecular weight of the IgG heavy chain was comparable to that of fully glycosylated intact rabbit IgG ( Figure 6 A, staining, 0 hours and IgG). In contrast, there was already a partial shift of th...

Embodiment 3

[0124] Example 3: EndoS is active in rabbits despite antibodies against the EndoS enzyme

[0125] Since EndoS was fully active at the second and third injections, it was intriguing to determine whether this was due to an absent or low level of immune response to the enzyme or to have specific antibodies against EndoS that did not interfere with enzymatic activity. Interested in. Since it is known that healthy individuals and those infected with S. pyogenes have antibodies against EndoS ( This is of particular interest since et al., 2004, J. Infect. Dis. 189:797-804).

[0126] To investigate this, purified rEndoS was separated on 10% SDS-PAGE and electroblotted onto PVDF cut into 1.5 mn strips. Strips were incubated with a 1:500 dilution of all serum samples from the first, second and third injections, followed by incubation with peroxidase-labeled goat anti-rabbit antibody (Pierce). Strips were developed with chemiluminescence as described above for lectin blots.

[012...

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 provides use of an EndoS polypeptide, or a polynucleotide encoding an EndoS polypeptide, in the manufacture of a medicament for the treatment or prevention of a disease or condition mediated by IgG antibodies.

Description

field of invention [0001] The present invention relates to methods for treating or preventing diseases or disorders mediated by IgG antibodies, such as autoimmune diseases, transplant rejection, post-surgical therapy, and acquired hemophilia. Background of the invention [0002] IgG is a heterotetramer comprising two heavy and two light chains held together by disulfide bonds, forming a three-protein domain separated by a flexible and protease-sensitive hinge region. Two identical Fab portions bind antigen while a separate Fc portion is responsible for effector functions, including binding and activation of complement factor CIq and Fc receptors on leukocytes. [0003] In addition to the polypeptide backbone, the Fc portion contains a conserved glycan linked to Asn-297 on each heavy chain. This oligosaccharide has a complex biantennary pattern with core fucose linked to the innermost N-acetylglucosamine (GlcNAc). These glycans are located at C H 2 domain (the second const...

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 Patents(China)
IPC IPC(8): A61K38/47A61K38/16A61P37/00
CPCA61K48/005A61K38/47A61P1/04A61P1/08A61P1/16A61P13/12A61P15/00A61P17/00A61P17/06A61P17/14A61P19/02A61P19/04A61P21/00A61P21/04A61P25/00A61P27/02A61P27/16A61P29/00A61P3/10A61P35/00A61P37/00A61P37/02A61P37/06A61P5/00A61P5/06A61P5/14A61P5/18A61P5/40A61P7/00A61P7/04A61P7/06A61P7/08A61P9/00A61P9/08
Inventor L·博乔克马迪亚斯·科林阿恩·奥尔森R·霍默达K·S·南达库玛乌纳·莎侬
Owner ハンサバイオファルマアクチボラゲット
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