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

Bifunctional antibodies and uses thereof

A bifunctional antibody and multipurpose technology, applied in the fields of antibodies, anti-inflammatory agents, digestive system, etc., can solve the problems of limited efficacy of monoclonal antibodies

Active Publication Date: 2016-02-03
叶才果
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Monoclonal antibodies targeting a single disease target have limited efficacy

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
  • Bifunctional antibodies and uses thereof
  • Bifunctional antibodies and uses thereof
  • Bifunctional antibodies and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Protein preparation of bifunctional antibodies BIAU003, BIAU022 and BIAU023 of the present invention

[0025] According to the heavy chain and light chain amino acid sequences of bifunctional antibodies BIAU003, BIAU022 and BIAU023, cDNAs of heavy chains and light chains of bifunctional antibodies BIAU003, BIAU022 and BIAU023 were artificially synthesized, respectively. All cDNAs are codon-optimized for expression in mammalian cells. The synthesized cDNAs were cloned into pcDNA3.1 plasmids respectively, and the plasmids were constructed correctly by sequencing. The sequenced plasmid was transformed into the TOP10 strain, and a single colony was picked and inoculated into 0.5 liters of LB liquid medium. When the OD600 was 0.8, the bacteria were collected by centrifugation, and the plasmid was extracted with a plasmid extraction kit (purchased from Qiagen). Co-transfect the plasmid containing the light chain of the bifunctional antibody BIAU003 identified corr...

Embodiment 2

[0026] Example 2: SPR determination of the antigen-binding ability of the bifunctional antibodies BIAU003, BIAU022 and BIAU023 of the present invention

[0027] The affinity and binding kinetics of p40 and TNF-α (purchased from GE) were analyzed by SPR. p40 or TNF-[alpha] was covalently attached to the chip via primary amines (carboxymethyl dextran-coated chip) using standard amine coupling chemistry and chips provided by GE. The recombinant fusion protein was covalently coupled to biotin using a biotin labeling kit (Pierce Company), and then flowed through an avidin-labeled SA chip (purchased from GE Company), so that the reaction value RU reached 450. Association and dissociation constants were measured by flowing antibodies in PBS buffer at concentrations of 0.01, 0.03, 0.09, 0.27 μΜ and a flow rate of 50 μl / min. Antigen-antibody binding kinetics were followed for 3 minutes and dissociation kinetics were followed for 10 minutes. The binding and dissociation curves were fi...

Embodiment 3

[0032] Example 3: Competition ELISA method to determine that the bifunctional antibodies BIAU003, BIAU022 and BIAU023 of the present invention compete with TNF-α receptors for binding to antigen TNF-α

[0033] Coat the ELISA plate with TNF-α-mFc, block with 1% BSA, mix different concentrations of antibodies BIAU003, BIAU022, BIAU023 with TNF-α receptor-hFc, incubate at 37°C, add the enzyme-labeled secondary antibody and incubate at 37°C 30 minutes. The absorbance at 450 nm was detected on a microplate reader. The results of the binding of the bifunctional antibody to the antigen TNF-α show that the bifunctional antibodies BIAU003, BIAU022, and BIAU023 of the present invention can all effectively compete with the TNF-α receptor for binding to the TNF-α protein, and the binding efficiency is dose-dependent. Through the analysis of the competition ELISA results of the combined bifunctional antibodies, the curve simulation bifunctional antibodies BIAU003, BIAU022, and BIAU023 ant...

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 three bifunctional antibodies, the three bifunctional antibodies can be specifically combined with human TNF-alpha, and also can be specifically combined with human P40. The three bifunctional antibodies all are capable of extremely effectively blocking combination of human IL-12 and IL-12 acceptor, and all are capable of extremely effectively blocking combination of human TNF-alpha and TNF-alpha acceptor. The three bifunctional antibodies are capable of simultaneously combining human TNF-alpha and human P40, and are capable of effectively inhibiting combination of TNF-alpha and IL-12 with responding acceptors, thereby respectively inhibiting secretion of inflammation cytokine and IFNgamma molecule. The three bifunctional antibodies are applicable to treat rheumatoid arthritis, chronic plaque psoriasis, moderate to severe Crohn's disease, moderate to severe ulcerative colitis, ankylosing spondylitis, psoriatic arthritis, moderate to severe multi-joint type juvenile idiopathic arthritis and Crohn's disease paediatrics patients.

Description

technical field [0001] The invention relates to an antibody, in particular to a bifunctional antibody capable of simultaneously blocking the binding of human TNF-α and TNF-α receptors and blocking the binding of human IL-12 and IL-12 receptors. Background technique [0002] Humira (Adalimumab) is a recombinant fully humanized tumor necrosis factor α monoclonal antibody expressed in Chinese hamster ovary cells. In addition to moderate-to-severe rheumatoid arthritis, FDA-approved indications include moderate-to-severe chronic plaque psoriasis, moderate-to-severe Crohn's disease, moderate-to-severe ulcerative colitis, ankylosing Therapeutic use in pediatric patients with spondylitis, psoriatic arthritis, moderate to severe polyarticular juvenile idiopathic arthritis, and Crohn's disease. In 2013, among the top ten best-selling branded drugs in the world, adalimumab ranked first with sales of US$10.66 billion and a growth rate of 15.0%. [0003] The monoclonal antibody Stelara...

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
IPC IPC(8): C07K16/46A61K39/395A61P29/00A61P19/02A61P17/06A61P1/04A61P1/00A61P25/00
Inventor 叶才果
Owner 叶才果
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