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

Immunoglobulin single variant antigen binding domain and specific construct thereof

An immunoglobulin and binding specificity technology, applied in the field of preparation of dual-specific ligands, can solve the problems of lack of light chain partners, limited therapeutic value, poor stability and qualitative properties, etc.

Inactive Publication Date: 2008-09-10
DORMANTIS LTD
View PDF15 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, these single domains have a short half-life in vivo, thus limiting their therapeutic value
[0011] It has been suggested that heavy chain variable domains of different specificities be joined together to produce bispecific antibody fragments (as described above), but this strategy has the disadvantage that the isolated antibody variable domains may have a domain that normally interacts with the light chain. Acting hydrophobic interface, which is exposed to solvent and can be sticky, allowing the single domain to bind to the hydrophobic surface
Furthermore, the combination of two or more distinct heavy chain variable domains lacking a light chain partner and possibly through a hydrophobic interface prevents them from binding one rather than two ligands that they would be able to bind independently
Moreover, in this case, the heavy chain variable region does not combine with the complementary light chain variable region, so the stability is poor and the folding is easy to open (Worn and Pluckthun, Biochemistry 37:13120-7 (1998))

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
  • Immunoglobulin single variant antigen binding domain and specific construct thereof
  • Immunoglobulin single variant antigen binding domain and specific construct thereof
  • Immunoglobulin single variant antigen binding domain and specific construct thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0324] Preparation of immunoglobulin-based multispecific ligands

[0325] Whether the dual-specific ligand of the present invention is based on the open or closed conformation in the expected configuration of the invention, it can be prepared according to the previously established technology applied in antibody engineering. Such as preparation of scFv, phage antibody and other engineered antibody molecules. Examples of techniques for preparing antibodies, particularly bispecific antibodies, are described in the following reviews and citations: Winter & Milstein, (1991) Nature 349:293-299; Plueckthun (1992) Immunological Reviews 130:151-188; Wright et al. , (1992) Crti.Rev.Immunol.12: 125-168; Holliger, P. & Winter, G. (1993) Curr.Op.Biotechn.4, 446-449; Carter, et al. (1995) J. Hematother.4, 463-470; Chester, K.A. & Hawkins, R.E. (1995) Trends Biotechn.13, 294-300; Hoogenboom, H.R. (1997) Nature Biotechnol.15, 125-126; Fearon, D. (1997) Nature Biotechnol.15, 618-619; Pliick...

Embodiment 1

[0483] Example 1: Screening of bispecific scFv antibody (K8) against human serum albumin (HSA) and β-galactosidase (β-gal)

[0484] This example illustrates a method for preparing bispecific antibodies against β-gal and HSA, selected for binding to β-gal and germline (mock) V H area connected to V κ Variable region repertoire, selected for ability to bind HSA and germline (mock) V κ area connected to V H variable region library. Then combine the selected V H HSA and Vκβ-gal variable regions, and antibodies were selected based on their ability to bind β-gal and HAS. HSA is a half-life enhancing protein found in human blood.

[0485] Four human phage antibody libraries used in this experiment.

[0486] Pool 1 Germline V κ / DVTV H 8.46×10 7

[0487] Pool 2 Germline V κ / NNKV H 9.64×10 7

[0488] Pool 3 Germline V H / DVTV κ 1.47×10 8

[0489] Pool 4 Germline V H / NNKV κ 1.45×10 8

[0490] All antibody libraries are based on a single human V H...

Embodiment 2

[0496] Example 2: Characterization of K8 Antibody Binding Performance

[0497] First, the binding properties of the K8 antibody were characterized by monoclonal phage ELISA. Use 100 μl of HSA and β-gal at a concentration of 10 μg / ml in PBS together with alkaline phosphatase (APS), bovine serum albumin (BSA), peanut agglutinin, lysozyme and cytochrome C (to check cross-reactivity ) to coat 96-well plates overnight at 4°C. Phagemids from K8 clones were rescued with KM13 as described by Harrison et al. (1996) and the phage-containing supernatants (50 [mu]l) were analyzed directly. A standard ELISA procedure (Hoogenboom et al., 1991) was then performed using an anti-M13-HRP conjugate to detect bound phage. When the absorption signal displayed on the surface of the phage is greater than 1.0, it is found that the bispecific K8 antibody can bind HAS and β-gal ( Figure 4 ). Strong binding to BSA was also observed ( Figure 4 ). Since HSA and BSA are 76% homologous at the amino ...

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 a dual-specific ligand comprising a first immunoglobulin variable domain having a first binding specificity and a complementary or non-complementary immunoglobulin variable domain having a second binding specificity.

Description

[0001] This application is a divisional application of an invention application with a filing date of June 30, 2003, a Chinese application number of 03820458.4, and an invention title of "antigen-binding region of a single variant of immunoglobulin and its specific construct". [0002] related application [0003] The present invention relates to dual specific ligands. The invention specifically provides a method for preparing a dual-specific ligand comprising a first immunoglobulin single variable domain that binds a first antigen or epitope, and a second immunoglobulin single variable domain that binds a second antigen or epitope. A single variable region of a globulin. More specifically, the present invention relates to dual specific ligands, the binding of which to at least one of a first and a second antigen or epitope prolongs the in vivo half-life of the ligand. The present invention describes open and closed conformational ligands comprising more than one binding speci...

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): C07K16/46C07K16/24C07K16/28C07K16/18C12N15/13C12N15/62C12N15/63A61K39/395A61P35/00A61P29/00G01N33/533C07K16/40C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12P21/08G01N21/78G01N33/535
CPCC07K16/18C07K16/40C07K16/468C07K2317/55C07K2317/622C07K2319/00
Inventor 格雷格·温特伊恩·汤姆林森奥尔加·伊格内托维奇露西·霍尔特埃琳娜·德安杰利斯菲利普·琼斯
Owner DORMANTIS LTD
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