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

Isolated antibody or antigen-binding fragment thereof and application of isolated antibody or antigen-binding fragment thereof in tumor therapy

A technology that combines fragments and antigens, applied in anti-tumor drugs, anti-receptors/cell surface antigens/cell surface determinant immunoglobulins, antibodies, etc., can solve problems such as lack of immunity

Active Publication Date: 2018-06-29
NANJING UMAB-BIOPHARMA CO LTD
View PDF8 Cites 14 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Interestingly, the mice in the treated group still had good immunity to re-inoculation with the same tumor, but not similar immunity to inoculation with different tumor cells [33]

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
  • Isolated antibody or antigen-binding fragment thereof and application of isolated antibody or antigen-binding fragment thereof in tumor therapy
  • Isolated antibody or antigen-binding fragment thereof and application of isolated antibody or antigen-binding fragment thereof in tumor therapy
  • Isolated antibody or antigen-binding fragment thereof and application of isolated antibody or antigen-binding fragment thereof in tumor therapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0203] Example 1: Obtaining of mouse anti-human OX40 activating monoclonal antibody

[0204] The inventors of the present application constructed a CHO cell line overexpressing OX40, used it to immunize mice, and performed hybridoma cell fusion. Through antigen-antibody binding experiments and cell function screening, a series of murine antibodies with strong ELISA binding activity and OX40 activation function were obtained. The sequences of the variable regions VH and VL of each antibody were obtained by sequencing, and human-mouse chimeric antibodies were designed and expressed accordingly. The heavy chain variable region was selected from the following sequences: SEQ ID NO: 1, SEQ ID One or more of NO:2, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11. The light chain variable region is selected from the following sequences: SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 12, SEQ ID NO: 13 and SEQ ID NO : One or more of 14.

[0205...

Embodiment 2

[0215] Embodiment 2: ELISA binding experiment

[0216] The protein solution with a concentration of 1 μg / mL was coated on a 96-well high-affinity plate at 100 μL / well, and shaken overnight at 4 °C. The next day, wash 3 times with 300 μL PBST (Tween20: 0.5‰), then block with 100 μL / well of 5% BSA / PBS for 2 hours, and shake at room temperature. Wash 3 times with 300 μL PBST. Prepare serial dilutions of samples in PBS. Add 100 μL / well to a 96-well plate and shake at room temperature for 1 hour. Wash 3 times with 300 μL PBST. Prepare the secondary antibody goat anti-human IgG HRP solution, add 100 μL / well to the 96-well plate, and shake at room temperature for 1 hour. Wash 4 times with 300 μL BST. Add 100μL / well TMB, develop color for 20min. Add 100 μL / well 0.6N H2SO4 to stop color development and detect OD450nm.

[0217] After testing, the results of figure 1 As shown, the ELISA binding EC50 of human-mouse chimeric antibodies MT01-L1 and MT01-L2 are 290.7ng / mL and 208.5ng...

Embodiment 3

[0218] Example 3: Binding to CHO-hOX40

[0219] Use PBS to prepare the OX40 antibody gradient, and prepare a final concentration of 10× working solution. Collect CHO-hOX40 cells, wash once with PBS, count, and dilute to 2*10^6 / ml cell suspension; add 10 μl OX40 antibody working solution to 100 μl cell suspension, and incubate at 4°C in the dark for 30 minutes; wash with PBS twice , add secondary antibody, incubate at 4°C in the dark for 30 min, wash once with PBS, suspend with 400 μl FACS buffer, and detect on the machine. Such as figure 2 As shown, the results show that MT01-L1 and MT01-L2 bind to human OX40, and their EC50 are 1.22 μg / mL and 2.42 μg / mL, respectively.

[0220] Similarly, the inventors of the present application tested the binding of antibodies to CHO cells expressing mouse OX40, see image 3 As a result of the binding experiment, it was found that neither MT01-L1 nor MT01-L2 combined with mouse OX40.

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 isolated antibody or an antigen-binding fragment thereof and application of the isolated antibody or the antigen-binding fragment thereof in tumor therapy. The isolated antibody or the antigen-binding fragment thereof can be used for preparing medicines for treating tumors.

Description

technical field [0001] The present invention relates to an antibody, in particular to an isolated antibody or an antigen-binding fragment thereof and its application in tumor therapy. Background technique [0002] Human OX40 is a 277aa protein, but due to its glycosylation at N146 and N160, its apparent molecular weight is about 50kD[1,2]. OX40 is a type I transmembrane protein, its extracellular part can bind to its natural ligand OX40L (CD252), and its intracellular part is coupled with multiple signaling pathways of T cell activation. [0003] Human OX40 is mainly expressed on activated T cells, including CD4, CD8, Th and Treg cells (reviewed in[3]). exist The expression of OX40 on T cells is very low, but its expression level is up-regulated after antigen-induced stimulation and reaches a peak within 12h to 5-6 days. Similarly, the expression of OX40L is also affected by the state of cell activation [3]. The expression of OX40L can be detected in APC cells 1-3 days ...

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): C07K16/28C12N15/13A61K39/395A61P35/00A61P31/12A61P31/20A61P31/22A61P31/18G01N33/68G01N33/574G01N33/569
CPCG01N33/56983G01N33/574G01N33/6872C07K16/2875A61K2039/505C07K2317/24C07K2317/622C07K2317/56C07K2317/55C07K2317/54C07K2317/569G01N2800/52G01N2800/24G01N33/6893C07K16/2878C07K2317/75C07K2317/76C07K2317/732C07K2317/74C07K2317/94C07K2317/565G01N33/68G01N2333/70578
Inventor 殷迅赵静
Owner NANJING UMAB-BIOPHARMA CO 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