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

Anti-HER3 antibodies and uses thereof

An antibody and expression vector technology, applied in the field of antibodies that bind to human HER3, which can solve problems such as incomplete characterization

Active Publication Date: 2014-09-24
ROCHE GLYCART AG
View PDF35 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Other splice variants have also been reported, but they have not been thoroughly characterized

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-HER3 antibodies and uses thereof
  • Anti-HER3 antibodies and uses thereof
  • Anti-HER3 antibodies and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0140] immunity

[0141] NMRI mice were immunized with hHER3-ECD (own use) and boosted with huHER3-ECD. The immune response was monitored by testing serum samples against HER1 / 2 / 3-ECD-ELISA. Splenocytes from mice with sufficient anti-HER3 immunoglobulin titers were frozen for later immortalization by fusion with the mouse myeloma cell line P3X63 Ag8.653. One fusion was done, and hybridoma supernatants were screened by HER1 / 2 / -ECD-ELISA that did not show cross-reactivity but combined with HER3-ECD, and anti-HER3 selective hybridomas were selected. Clonal related hybridomas were sorted by single-cell FACS. Single cell clones from different hybridomas were cultured in vitro to produce antibodies in tissue culture for characterization. Antibodies were selected by assaying their ability to inhibit HER3 phosphorylation, AKT phosphorylation, and tumor cell proliferation of MDA-MB-175 cells (see Examples below). From the obtained antibodies, one antibody was further humanized to g...

Embodiment 2

[0146] binding assay

[0147] a) Antigen-specific ELISA for binding to human HER3 ECD

[0148] Soluble human HER3 ectodomain fused to streptavidin binding protein (SBP) was captured on streptavidin plates. In order to define the optimal binding of the antibody to SPB-CDCP1, pure and step-wise diluted HEK293 supernatant (in BSA / IMDM buffer: 100 mg / ml BSA fraction V, Roche 10735078001, cultured in Iscove's modified Dulbeccos 384-well polystyrene plates (NUNC, streptavidin-coated) supplied by MicroCoat, Bernried, Germany (ID-No. 1734776-001). Using a mouse calibration curve for the chimeric 205 antibody, the optimal dilution factor of HEK293 supernatant relative to the streptavidin binding capacity of the microtiter plate was identified. For standard coating, HEK293 supernatants containing SBP-HER3 were diluted (between 1:15 and 1:40) and incubated overnight at 2-80C (25 μl per well). Vigorous washing of the microtiter plate is necessary to remove remaining unbound SBP-HER3. ...

Embodiment 3

[0159] a) Inhibition of HER3 phosphorylation in MCF7, FaDu and Mel-Juso cells

[0160] Assays were performed in MCF7 and FaDu cells according to the following protocol: Cells were seeded at 500,000 cells / well in poly-D-lysine coated 6-well plates in RPMI1640 medium with 10% FCS. Incubate for 24 hours. Media was removed by aspiration and incubated overnight in 500 μl / well RPMI1640 with 0.5% FCS. Add 500 μl of antibody in RPMI 1640 with 0.5% FCS. Incubate for 1 hour. Add HRG-1b (final concentration 500 ng / ml) for 10 minutes. To lyse the cells, the medium was removed and 80 μl of ice-cold Triton-X-100 cell lysis buffer was added and incubated on ice for 5 minutes. After transferring the lysate into a 1.5 ml reaction tube and centrifuging at 14000 rpm for 15 minutes at 4°C, the supernatant was transferred into a fresh reaction tube. Samples containing equal amounts of protein in SDS loading buffer were resolved on SDS PAGE and blotted to nitrocellulose membranes by using semi...

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 relates to antibodies binding to human HER3 (anti-HER3 antibody), methods for their production, pharmaceutical compositions containing said antibodies, and uses thereof.

Description

[0001] The present invention relates to an antibody that binds to human HER3 (anti-HER3 antibody), a method for its production, a pharmaceutical composition containing the antibody, and uses thereof. Background of the invention [0002] Human HER3 (ErbB-3, ERBB3, c-erbB-3, c-erbB3, receptor tyrosine protein kinase erbB-3, SEQ ID NO:17) encodes the epidermal growth factor receptor (EGFR) receptor tyrosine A member of the kinase family that also includes HER1 (also known as EGFR), HER2, and HER4 (Kraus, M.H. et al., PNAS 86(1989) 9193-9197; Plowman, G.D. et al., PNAS 87(1990) 4905- 4909; Kraus, M.H. et al., PNAS 90 (1993) 2900-2904). Like the prototypical EGFR, the transmembrane receptor HER3 consists of an extracellular ligand-binding domain (ECD), a dimerization domain within the ECD, a transmembrane domain, an intracellular protein tyrosine kinase domain (TKD), and a C terminal phosphorylation domain. This membrane-bound protein has a HER3 heregulin (HRG) binding domain wit...

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): C07K16/28C07K16/32A61K39/395A61P35/00
CPCC07K2317/92C07K16/32C07K2317/732A61K2039/505C07K2317/73A61P35/00C07K16/28A61K39/395C07K16/40C07K2317/76C07K2317/24C07K2317/41C07K2317/565
Inventor B.伯森梅尔N.迪默迪斯T.弗里斯G.乔治斯I.科尔姆H-W.克雷尔V.利夫克E.莫斯纳
Owner ROCHE GLYCART AG
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