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

Human antibodies that bind human il-12 and methods for producing

a technology of human il-12 and antibodies, which is applied in the field of human antibodies, can solve the problems of not being described, eliciting unwanted immune reactions, and limited use of murine il-12 antibodies in vivo, and achieves the effect of improving activity

Inactive Publication Date: 2008-12-11
ABBVIE DEUTSHLAND GMBH & CO KG
View PDF3 Cites 57 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The selectively mutated human antibodies demonstrate improved binding specificity and neutralization of IL-12, with dissociation rates and inhibitory capacities superior to previous antibodies, effectively inhibiting IL-12 activity in autoimmune and inflammatory conditions without triggering adverse immune reactions.

Problems solved by technology

These murine IL-12 antibodies are limited for their use in vivo due to problems associated with administration of mouse antibodies to humans, such as short serum half life, an inability to trigger certain human effector functions and elicitation of an unwanted immune response against the mouse antibody in a human (the “human anti-mouse antibody” (HAMA) reaction).
However, because these chimeric and humanized antibodies still retain some murine sequences, they still may elicit an unwanted immune reaction, the human anti-chimeric antibody (HACA) reaction, especially when administered for prolonged periods.
However, such antibodies have not been described in the art and, therefore are still needed.

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
  • Human antibodies that bind human il-12 and methods for producing
  • Human antibodies that bind human il-12 and methods for producing
  • Human antibodies that bind human il-12 and methods for producing

Examples

Experimental program
Comparison scheme
Effect test

example 1

Isolation of Anti-IL-12 Antibodies

A. Screening for IL-12 Binding Antibodies

[0615]Antibodies to hIL-12 were isolated by screening three separate scFv phage display libraries prepared using human VL and VH cDNAs from mRNA derived from human tonsils (referred to as scFv 1), tonsil and peripheral blood lymphocytes (PBL) (referred to as scFv 2), and bone marrow-derived lymphocytes (referred to as BMDL). Construction of the library and methods for selection are described in Vaughan et al. (1996) Nature Biotech. 14: 309-314.

[0616]The libraries were screened using the antigens, human IL-12 p70 subunit, human IL-12 p40 subunit, chimaeric IL-12 (mouse p40 / human p35), mouse IL-12, biotinylated human IL-12 and biotinylated chimaeric IL-12. IL-12 specific antibodies were selected by coating the antigen onto immunotubes using standard procedures (Marks et al., (1991) J. Mol. Biol. 222: 581-597). The scFv library 2 was screened using either IL-12, or biotinylated-IL-12, and generated a significant...

example 2

Mutation of Y61 at Hypermutation and Contact Positions

[0631]Typically selection of recombinant antibodies with improved affinities can be carried out using phage display methods. This is accomplished by randomly mutating combinations of CDR residues to generate large libraries containing single-chain antibodies of different sequences. Typically, antibodies with improved affinities are selected based on their ability to reach an equilibrium in an antibody-antigen reaction. However, when Y61 scFV was expressed on phage surface and incubated with IL-12, selection conditions could not be found that would allow the system to reach normal antibody-antigen equilibrium. The scFV-phage remained bound to IL-12, presumably due to a non-specific interaction, since purified Y61 scFv exhibits normal dissociation kinetics. Since the usual methods of phage-display affinity maturation to Y61 (i.e. library generation and selections by mutagenesis of multiple CDR residues) could not be utilized, a new...

example 3

Functional Activity of Anti-hIL-12 Antibodies

[0639]To examine the functional activity of the human anti-human IL-12 antibodies of the invention, the antibodies were used in several assays that measure the ability of an antibody to inhibit IL-12 activity.

A. Preparation of Human PHA-Activated Lymphoblasts

[0640]Human peripheral blood mononuclear cells (PBMC) were isolated from a leukopac collected from a healthy donor by Ficoll-Hypaque gradient centrifugation for 45 minutes at 1500 rpm as described in Current Protocols in Immunology, Unit 7.1. PBMC at the interface of the aqueous blood solution and the lymphocyte separation medium were collected and washed three times with phosphate-buffered saline (PBS) by centrifugation for 15 minutes at 1500 rpm to remove Ficoll-Paque particles.

[0641]The PBMC were then activated to form lymphoblasts as described in Current Protocols in Immunology, Unit 6.16. The washed PBMC were resuspended at 0.5−1×106 cells / ml in RPMI complete medium (RPMI 1640 me...

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

PropertyMeasurementUnit
autoimmune disorderaaaaaaaaaa
surface plasmon resonanceaaaaaaaaaa
affinityaaaaaaaaaa
Login to View More

Abstract

Human antibodies, preferably recombinant human antibodies, that specifically bind to human interleukin-12 (hIL-12) are disclosed. Preferred antibodies have high affinity for hIL-12 and neutralize hIL-12 activity in vitro and in vivo. An antibody of the invention can be a full-length antibody or an antigen-binding portion thereof. The antibodies, or antibody portions, of the invention are useful for detecting hIL-12 and for inhibiting hIL-12 activity, e.g., in a human subject suffering from a disorder in which hIL-12 activity is detrimental. Nucleic acids, vectors and host cells for expressing the recombinant human antibodies of the invention, and methods of synthesizing the recombinant human antibodies, are also encompassed by the invention.

Description

RELATED APPLICATIONS[0001]This application is a divisional of U.S. patent application Ser. No. 09 / 534,717, filed Mar. 24, 2000, which is a non-provisional application claiming priority to U.S. provisional application Ser. No. 60 / 126,603, filed Mar. 25, 1999, the contents of which are hereby incorporated by reference.BACKGROUND OF THE INVENTION[0002]Human interleukin 12 (IL-12) has recently been characterized as a cytokine with a unique structure and pleiotropic effects (Kobayashi, et al. (1989) J. Exp Med. 170:827-845; Seder, et al. (1993) Proc. Natl. Acad. Sci. 90:10188-10192; Ling, et al. (1995) J. Exp Med. 154:116-127; Podlaski, et al. (1992) Arch. Biochem. Biophys. 294:230-237). IL-12 plays a critical role in the pathology associated with several diseases involving immune and inflammatory responses. A review of IL-12, its biological activities, and its role in disease can be found in Gately et al. (1998) Ann. Rev. Immunol. 16: 495-521.[0003]Structurally, IL-12 is a heterodimeric...

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(United States)
IPC IPC(8): A61K39/395A61P37/00C07K16/24
CPCA61K2039/505C07K2317/94C07K2317/21C07K2317/565C07K2317/73A61K39/39591A61K45/06C07K2317/76A61K39/3955C07K16/00C07K16/18C07K14/5434A61K39/00A61K38/208Y10S514/885Y10S514/863Y10S514/886A61K9/0019A61K47/20A61K47/22A61K47/26C07K16/244A61P1/00A61P1/04A61P1/16A61P11/06A61P13/12A61P17/06A61P19/02A61P25/00A61P25/16A61P25/28A61P37/00A61P9/10A61P3/10
Inventor SALFELD, JOCHENROGUSKA, MICHAELPASKIND, MICHAELBANERJEE, SUBHASHISTRACEY, DANIELWHITE, MICHAELKAYMAKCALAN, ZEHRALABKOVSKY, BORISSAKORAFAS, PAULVELDMAN, GEERTRUIDA M.VENTURINI, AMYWIDOM, ANGELAFRIEDRICH, STUARTWARNER, NICHOLAS W.MYLES, ANGELAELVIN, JOHN GAWAINDUNCAN, ALEXANDER ROBERTDERBYSHIRE, ELAINE JOYCARMEN, SARASMITH, STEPHENHOLTET, THOR LASDU FOU, SARAH LEILA
Owner ABBVIE DEUTSHLAND GMBH & CO KG
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