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

Compositions and methods of use for therapeutic low density lipoprotein-related protein 6 (LPR6) multivalent antibodies

A multivalent antibody, antibody technology, applied in the direction of antibody mimics/scaffolds, antibodies, drug combinations, etc., can solve the problems of loss of antibody activity, low efficiency, and troublesome purification of antibodies.

Inactive Publication Date: 2013-08-07
NOVARTIS AG
View PDF160 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, purification of antibodies from non-functional species such as homodimers and mispaired heterodimers of unrelated Ig light and heavy chains produced by hybrid hybridomas is cumbersome and inefficient
[0009] Chemical cross-linking of two IgG or fragments thereof is also inefficient and may result in loss of antibody activity (Zhu et al. (1994) Cancer Lett. 86:127-34)
Multimeric aggregates caused by chemical conjugation lead to low yields (Cao et al. (1998) Bioconj. Chem. 9:635-44)

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
  • Compositions and methods of use for therapeutic low density lipoprotein-related protein 6 (LPR6) multivalent antibodies
  • Compositions and methods of use for therapeutic low density lipoprotein-related protein 6 (LPR6) multivalent antibodies
  • Compositions and methods of use for therapeutic low density lipoprotein-related protein 6 (LPR6) multivalent antibodies

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0636] Example 1: Determination of specific binding of anti-LRP6 antibodies to endogenous LRP6 by FACS

[0637] Detection of endogenous cell surface expression of LRP6 was examined on a number of tumor cells using anti-LRP6 antibodies and FACS analysis. as in Figure 1A As shown in , PA1 cells express LRP5 and LRP6 mRNA, while U266 and Daudi cells do not express LRP6 mRNA. PA1 cells, but not U266 and Daudi cells showed significant staining by Propeller 1 anti-LRP6 IgG. More importantly, U266 cells were not stained by anti-LRP6 antibody, although they expressed LRP6, indicating the specificity of anti-LRP6 antibody. Furthermore, when endogenous LRP6 was depleted using LRP6 shRNA, the staining of PA1 cells by anti-LRP6 antibody was significantly reduced, further indicating the specificity of LRP6 antibody.

[0638] In other studies, the specificity of Prop1 and Prop3 for LRP6 was further confirmed by shRNA knockdown of LRP6 in PA1 cells (see Figure 1B ). Knockdown was achi...

Embodiment 2

[0639] Example 2: Differential inhibition of Wnt1 and Wnt3a reporter gene assays by Propeller 1 and Propeller 3 anti-LRP6 Fabs

[0640] Different anti-LRP6 Fabs were tested in an in vitro Wnt reporter assay. Wnt1 or Wnt3A ligands (reporter gene assay) were transiently expressed in HEK293T / 17STF cells and treated with different concentrations of anti-LRP6 Fab fragments. STF assays were performed using the method described by Huang et al. (2009), Nature; 461 :614-20 (epublished September 16, 2009). as in Figure 2A As seen in Propeller 1 anti-LRP6 Fabs (MOR08168, MOR08545, MOR06706) specifically reduced Wnt1-dependent signaling without much effect on Wnt3A-dependent signaling. Conversely, as in Figure 2B As shown in Propeller 3 anti-LRP6 Fabs (MOR06475, MOR08193, MOR08473) specifically decreased Wnt3-dependent signal transduction without significant effect on Wnt1-dependent signal transduction. The results indicated that Wnt1 and Wnt3A activities were blocked separately by ...

Embodiment 3

[0641] Example 3: Binding of anti-LRP6 antibodies to LRP6 from different species

[0642] To show cross-reactivity, cells expressing endogenous LRP of human (HEK293T / 17) and mouse origin (NIH 3T3) or transiently transfected HEK293 / T17 cells expressing macaque LRP6 were treated as described above and subjected to Flow Cytometry. image 3 The findings were summarized and shown that all anti-LRP6 antibodies bound human, mouse and macaque LRP6.

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 disclosure relates to an antibody or antigen binding fragment having at least two receptor binding domains for two different binding sites of LRP6 and compositions and methods of use thereof.

Description

[0001] related application [0002] This application claims priority to US Provisional Application Serial No. 61 / 331,993, filed May 6, 2010, the contents of which are hereby incorporated by reference in their entirety. field of invention [0003] The present invention relates to multivalent antibodies comprising at least two receptor binding domains directed against two different binding sites on LRP6. More specifically, the present invention relates to multivalent antibodies that are LRP6 antagonists. Background of the invention [0004] The Wnt / β-catenin pathway regulates a variety of biological processes during development and tissue homeostasis by regulating the protein stability of β-catenin (Clevers et al., (2006) Cell 127:469-480; and Logan et al. , (2004) Annu. Rev Cell Dev. Biol 20:781-810). In the absence of Wnt signaling, cytoplasmic β-catenin binds to the β-catenin destruction complex, which contains a variety of proteins, including adenomatous polyposis coli (...

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): C07K16/28A61K39/395A61P35/00C07K16/46C12N15/113
CPCC07K2317/31C07K2317/34C07K2319/00C07K2317/21C07K2317/92C07K2317/94C07K2317/55C07K16/2863C07K2317/35C07K2317/33C07K2317/622C07K2317/64A61K2039/505C07K2317/76A61P1/04A61P1/16A61P11/00A61P13/08A61P13/10A61P15/00A61P17/00A61P19/00A61P21/00A61P35/00A61P35/02A61P43/00C07K16/28A61K39/395C07K16/46C07K16/468A61K39/3955
Inventor 丛枫S·埃滕伯格D·詹金斯雷鸣A·洛K·文森特周立
Owner NOVARTIS 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