Check patentability & draft patents in minutes with Patsnap Eureka AI!

Antibody specifically binding to CD276 or antigen binding fragment thereof, and preparation method and application thereof

A technology of CD276 and binding fragments, applied in botany equipment and methods, biochemical equipment and methods, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve the problem of losing curative effect and achieve good results safety effect

Active Publication Date: 2021-11-09
HRAIN BIOTECHNOLOGY CO LTD
View PDF9 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] As an important part of CAR, single-chain antibodies are currently traditionally used in mouse-derived antibodies. Due to the heterogeneity of mouse antibodies, human anti-mouse antibody reactions ( Human anti-mouse antibody reaction , HAMA), causing CAR-T to be quickly cleared in the circulatory system and lose its 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
  • Antibody specifically binding to CD276 or antigen binding fragment thereof, and preparation method and application thereof
  • Antibody specifically binding to CD276 or antigen binding fragment thereof, and preparation method and application thereof
  • Antibody specifically binding to CD276 or antigen binding fragment thereof, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] Example 1: Construction and eukaryotic expression of recombinant human CD276 protein expression vector.

[0103] 1. Synthesis of the gene sequence from the 29th to 466th amino acid interval of CD276 and the construction of the protein expression vector.

[0104] Import the 29th to 466th amino acid sequence of CD276 (uniprot accession No: Q5ZPR3-1) into the online codon optimization tool (http: / / www.jcat.de / #opennewwindow) to obtain the codon-optimized nucleic acid sequence, Then the gene sequence is obtained by chemical synthesis, and the nucleic acid sequence of avi-tag and 6×his-tag is added to the 3' end of the gene sequence, and the fusion gene sequence encodes the amino acid sequence shown in SEQ ID NO:39, SEQ ID The nucleotide sequence shown in NO:40. Through molecular cloning, the spliced ​​product was cloned into pCDNA3.1 (Thermo) with TaKaRa seamless cloning kit to obtain an expression vector.

[0105] 22. Expression, purification and activity identification ...

Embodiment 2

[0110] Example 2: Preparation of anti-human CD276 human antibody.

[0111] 1. Construct a human natural antibody phage display library.

[0112] The human natural antibody phage display library is constructed by a two-step method, that is, the light chain gene and heavy chain gene of the human natural antibody are connected to the phage display vector in two steps.

[0113] Using human immunoglobulin κ chain variable region forward primer (VK-mix-F) and light chain constant region reverse primer (CK-mix-R) to amplify human immunoglobulin κ chain gene by PCR using human PBMC cDNA as template , The PCR reaction conditions are as follows: 98°C pre-denaturation for 1 minute, followed by temperature cycling, 98°C denaturation for 30 seconds, 58°C annealing for 30 seconds, 72°C extension for 1 minute, 30 cycles, and 72°C final extension for 10 minutes. After the PCR product was subjected to 1% agarose gel electrophoresis, the κ chain gene fragment of about 750 bp was recovered with...

Embodiment 3

[0143] Example 3: Preparation of retrovirus stock solution containing anti-human CD276 chimeric antigen receptor element.

[0144] 1. Preparation of chimeric antigen receptor targeting human CD276 antigen.

[0145] Gene synthesis or cloning of single-chain antibody scFv containing anti-human CD276 antigen, chimeric antigen receptor sequence of hinge region, transmembrane region and intracellular signal segment, its structure is as follows Figure 5 shown. According to the difference of loaded scFv and intracellular signal, the chimeric antigen receptors were named 1C8-BBz, 1H12-BBz, 3C1-BBz and 3E5-BBz respectively, and their nucleotide sequences were SEQ ID NO:34, SEQ ID Shown in NO:35, SEQ ID NO:36 and SEQ ID NO:37. At the same time, the scFv of the known CD276 murine antibody (clone number: 376.96) reported in the literature was selected to construct a chimeric antigen receptor as a control, named 376.96-BBz, and its nucleotide sequence is shown in SEQ ID NO: 38, sequence...

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 discloses an antibody specifically binding to CD276 or an antigen binding fragment thereof, and a preparation method and application thereof, and relates to the technical field of biological immunotherapy. The antibody has good safety and a therapeutic effect of targeting CD276, and can be used for preparing immune effector cells targeting CD276, thereby providing a treatment or improvement way for diseases related to CD276 expression.

Description

technical field [0001] The present invention relates to the technical field of biological immunotherapy, in particular to an antibody specifically binding to CD276 or an antigen-binding fragment thereof and a preparation method and application thereof. Background technique [0002] CD276, also known as B7-H3, is an important immune checkpoint member of the B7 and CD28 families. It belongs to type I transmembrane protein. The extracellular region contains two pairs of identical immunoglobulin variable regions and constant regions, and the intracellular region is very short , with no clear signaling motif. Its expression at the mRNA level is relatively extensive, but its protein expression is relatively limited to non-immune cells such as resting fibroblasts, endothelial cells, osteoblasts, and amniotic fluid stem cells, as well as the surface of induced antigen-presenting cells and NK cells. [0003] Studies have shown that the expression of CD276 is abnormally elevated in a...

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): C07K16/28C07K19/00C12N15/13C12N5/10C12N15/867A61K39/395A61K39/00A61P35/00
CPCC07K16/2827C07K14/7051C12N5/0636C12N15/86A61K39/001129A61P35/00C07K2317/56C07K2317/565C07K2319/02C07K2317/92C07K2319/03C07K2319/33C12N2510/00C12N2730/00043
Inventor 胡红明彭涛黄慧
Owner HRAIN BIOTECHNOLOGY CO LTD
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com