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

Anti-PD-1 (programmed cell death-1) humanized antibody

A PD-1, human antibody technology, applied in the direction of antibodies, anti-animal/human immunoglobulins, anti-tumor drugs, etc., can solve the problem of only 10% single drug effective rate, achieve obvious affinity, inhibit ligand - effect of receptor binding

Active Publication Date: 2015-04-01
SHANGHAI HENLIUS BIOTECH INC
View PDF5 Cites 29 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The effective rate of the drug alone is only 10%

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-PD-1 (programmed cell death-1) humanized antibody
  • Anti-PD-1 (programmed cell death-1) humanized antibody
  • Anti-PD-1 (programmed cell death-1) humanized antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Expression and purification of human PD-1 extracellular domain recombinant protein PD-1-His

[0042] Synthetic human PD-1-AP-His fusion gene (synthesized by Nanjing GenScript Biotechnology Co., Ltd.), wherein the PD-1 gene sequence is SEQ ID NO: 5, the AP-His gene sequence is SEQ ID NO: 6, two The connection of the segment sequences is the PD-1-AP-His fusion gene. For the construction of the PD-1-His recombinant protein expression vector, the forward and reverse primers are designed as follows:

[0043] Forward primer:

[0044] 5'-ATCCGCCGGCAAGCCGCCACCATGCAGATCCCACAGGCGCCCTGG-3' (SEQ ID NO: 7);

[0045] Reverse primer:

[0046] 5'-TCACTACGTATCAGTGGTGGTGGTGGTGGTGTTGGAACTGGCCGGCTGGCCTGG-3'(SEQ ID NO:8), used to amplify the gene encoding PD-1-His fusion protein, and introduce restriction enzyme sites Ngo MIV and Sna BI at the upstream and downstream of the gene respectively , PCR conditions are 95°C for 30s, 60°C for 30s, 72°C for 30s, 35 cycles, 72°C extension for 10mi...

Embodiment 2

[0049] Expression and purification of human PD-1 extracellular region recombinant protein PD-1-AP-His

[0050] Using the PD-1-AP-His fusion gene synthesized in Example 1 as a template, the forward and reverse primers were designed as: 5'-ATCCGCCGGCAAGCCGCCACCATGCAGATCCCACAGGCGCCCTGG-3'(SEQ ID NO:7) and 5'-TCACTACGTATCAGTGGTGGTGGTGGTGGTGGCCTGAACC-3 '(SEQ ID NO: 9) is used to amplify the gene encoding the PD-1-AP-His fusion protein, introducing restriction endonuclease sites Ngo MIV and Sna BI at the upstream and downstream of the gene, and the PCR conditions are 95 30 s at ℃, 30 s at 60 ℃, 60 s at 72 ℃, 35 cycles, extension at 72 ℃ for 10 min, the DNA polymerase is the product of TAKARA company.

[0051] After the PCR product was recovered by agarose gel (the recovery kit is a product of Omega bio-tek), after purification, restriction endonucleases Ngo MIV and Sna BI were added for digestion, and the digested product was purified by a DNA recovery reagent (Omega bio-tek The pr...

Embodiment 3

[0054] Construction and expression of anti-PD-1 human antibody eukaryotic expression vector

[0055] The amino acid sequence of the antibody heavy chain variable region is SEQ ID NO: 2, and the coding DNA sequence used in this example is SEQ ID NO: 1;

[0056] The amino acid sequence of the light chain variable region of the antibody is SEQ ID NO: 4, and the coding DNA sequence used in this example is SEQ ID NO: 3; Synthetic Technology Co., Ltd.)

[0057] PCR reaction is used to amplify the coding DNA sequences of the above-mentioned light chain variable region and heavy chain variable region, and introduce appropriate enzyme cutting sites at both ends of the gene of the antibody heavy chain variable region and light chain variable region, wherein Ngo MIV and SnaBI restriction sites were introduced at the 5' end and 3' end of the antibody heavy chain variable region gene respectively, and the forward and reverse primer sequences were: 5'-ACCTGCCGGCAAGCCGCCACCATGGGTTGGAGCCTCAT...

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 relates to the technical field of biology, and in particular relates to an anti-PD-1 humanized antibody. The anti-PD-1 humanized antibody is characterized in that the amino acid sequence of a variable region of a heavy chain of the antibody is shown as SEQ ID NO: 2 or is a conserved variation sequence; the amino acid sequence of a variable region of a light chain of the antibody is shown as SEQ ID NO: 4 or is a conserved variation sequence. The anti-PD-1 humanized antibody shows relatively high expression quantity in mammalian cells, and is obvious in affinity with PD-1 and able to inhibit the combination of a ligand and a receptor.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to an anti-PD-1 human antibody. Background technique [0002] The activation of T cells is an important step in the functioning of the immune system, which not only requires the first signal provided by antigen-presenting cells (APC), but also is regulated by co-stimulatory signals (second signals). Co-stimulatory signal is a kind of membrane protein on the cell surface, which plays a role in co-regulating the proliferation, activation and differentiation of T cells. [0003] Programmed death receptor 1 (programmed death 1, PD-1), also known as CD279, is an important immunosuppressive molecule, belonging to the CD28 family member of the immunoglobulin superfamily, and a type I transmembrane with a molecular weight of 50-55kD glycoprotein. PD-1 is persistently expressed on the surface of T cells, B cells, natural killer cells, monocytes and myeloid cells. PD-1 has two known li...

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): C07K16/28C12N15/13C12N15/63C12P21/08A61K39/395A61P35/00
Inventor 肖辉郎国竣姜伟东
Owner SHANGHAI HENLIUS BIOTECH INC
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