Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Anti-human IL-33 monoclonal antibody and application thereof

A monoclonal antibody, IL-33 technology, applied in applications, antibodies, anti-animal/human immunoglobulins, etc., can solve the high adverse reaction rate, unclear clinical application prospects, and anti-IL-33 antibodies need further research And other issues

Active Publication Date: 2021-06-18
MABWELL (SHANGHAI) BIOSCIENCE CO LTD
View PDF6 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Although the mechanism is clear and the research direction is clear, due to many uncontrollable factors in the development of antibody drugs, anti-IL-33 antibodies still need to be further studied as clinical drugs
At present, the fastest progress in clinical trials is Itepekimab, a fully human anti-IL-33 monoclonal antibody developed by Sanofi and Regeneron, but its clinical application prospects in asthma, atopic dermatitis, and chronic obstructive pneumonia are still unclear
In addition, the fully human antibody Itepekimab has a high rate of adverse reactions clinically, and the incidence of adverse events (aes) of Itepekimab alone is 61.6%
Other studies on anti-IL-33 antibody drugs have insufficient effects in terms of humanization degree, affinity strength, and cytokine secretion

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-human IL-33 monoclonal antibody and application thereof
  • Anti-human IL-33 monoclonal antibody and application thereof
  • Anti-human IL-33 monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1: Preparation of anti-human IL-33 antibody mouse hybridoma cells

[0083] Human interleukin 33 (Ser112-Thr270) antigen was purchased from Novoprotein Technology Co., Ltd. (Novoprotein, product number C091), and 8-week-old Balb / c mice were selected as immunized animals, and blood was collected before immunization as a negative control. Human IL33 protein was emulsified in Freund's adjuvant and then subcutaneously injected for immunization three times with an interval of 2 weeks each time. Blood was collected from the tail vein one week after the third immunization to measure the antibody titer of the serum and select mice with high titer Sprint immunizations were performed 3 days prior to fusion.

[0084] Spleen cells and lymphocytes were aseptically separated from the spleen and lymph nodes of the immunized mice, fused with SP20 myeloma cells in good condition by electrofusion and PEG, and positive hybridomas were screened within 8 to 10 days. The hybridoma ce...

Embodiment 2

[0085] Example 2: The binding activity of anti-human IL33 monoclonal antibody secreted by hybridoma cell lines to human IL33

[0086] Take the supernatant of hybridoma cells 8-10 days after fusion, and use ELISA method to screen positive hybridoma mother clones that can secrete anti-human IL33 antibody.

[0087] The specific method is as follows: 0.5ug / ml human IL-33 protein, 30μl / well, coated 384-well plate (greiner bio-one 781097) and incubated overnight at 4°C; the next day, the plate was washed with PBST, and 60μl of IL-33 was added to the sample well Contain 1% BSA in PBST, block at 37°C for 2 hours; then wash the plate with PBST, and dry the water in the wells. Add 20 µl of the sample to be tested into the well plate and incubate at room temperature for 1 h. Then, after washing the plate with PBST, 30ul of 1:30000 diluted secondary antibody goat anti-mouse IgG coupled to horseradish peroxidase (Jackson, cat#: 115-035-164) was added to each well, and incubated at room te...

Embodiment 3

[0089] Example 3: Murine anti-human IL33 antibody blocks the binding of IL-33 to human ST2

[0090] Transfer the positive clones that bind to human IL33 protein from the 384-well plate to the 96-well plate, and use the ELISA method for blocking experiments. The specific method is as follows: coat the 96-well plate with hST2-hFc protein (Acro, #IL1-H5250), 1ug / ml, 50ul per well, incubated overnight at 4°C; the next day, wash the plate 3 times with PBST, add 200µl of PBST containing 1% BSA to the sample wells, block at 37°C for 2h; then wash the plate with PBST, and spin dry the wells Medium moisture. Add 50ul of pre-incubated samples (biotin hIL-33 protein and hybridoma clone supernatant) to the well plate and incubate at room temperature for 1 h. Then, after washing the plate with PBST, 50ul of 1:5000 diluted secondary antibody StrepTactin-HRP Conjugate (Bio-rad, #1610380) was added to each well, and incubated at room temperature for 1h. Wash the plate with PBST, add 50µl TM...

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 an anti-human IL-33 monoclonal antibody and application thereof. By taking recombinant human IL-33 as an immunogen, a murine anti-human IL-33 monoclonal antibody is prepared through a hybridoma technique; binding activity analysis is carried out on the murine antibody, blocking activity analysis is carried out on binding of IL-33 and ST2, sequencing is carried out, a human-mouse chimeric anti-human IL-33 monoclonal antibody is constructed, and kinetic parameters of the chimeric antibody are detected. On the basis of performance analysis of the murine antibody and the chimeric antibody, a humanized anti-human IL-33 monoclonal antibody is constructed by adopting a CDRs transplantation technology and a complementary determining region (CDR) mutation design, kinetic parameters and affinity of the humanized antibody are detected, and IL33-induced cytokine release is blocked. The anti-human IL-33 monoclonal antibody has relatively high affinity to IL-33, can effectively block binding of IL-33 and a receptor ST2 thereof, inhibits immune response related to an IL-33 / ST2 pathway, and has a good application prospect in treatment of asthma, inflammatory diseases, diabetes and autoimmune diseases.

Description

technical field [0001] The present invention belongs to the field of antibody engineering, in particular to an anti-human IL-33 monoclonal antibody and its application, in particular to a monoclonal antibody that blocks the binding of human IL-33 to ST2 and inhibits the secretion of cytokines induced by IL-33. Cloned antibodies and their applications. Background technique [0002] Interleukin 33 (IL-33) is a member of the interleukin 1 (IL-1) family, its gene is located on chromosome 9 (9p24. There is a helix-turn-helix structural pattern at the amino-terminus, which contains chromatin binding regions and nuclear localization sequences, and is a chromatin-related factor with transcriptional repression function; its carboxyl-terminus has a unique domain of IL-1 molecules, which is The binding site of IL-33 to its receptor. IL-33 is often expressed in barrier tissue cells, such as skin, intestine, lung, etc. When these cells are damaged, the full-length IL-33 (1-270) is extr...

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): C07K16/24C12N15/13C12P21/08A61K39/395A61P37/08A61P11/06A61P37/02A61P19/02A61P29/00A61P25/00A61P9/00A61P9/04
CPCA61P9/00A61P9/04A61P11/06A61P19/02A61P25/00A61P29/00A61P37/02A61P37/08C07K16/244C07K2317/24C07K2317/56C07K2317/565C07K2317/76C07K2317/92
Inventor 王晋邓小芳吴健徐晓红朱戬蔺利娟
Owner MABWELL (SHANGHAI) BIOSCIENCE CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
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