Anti-CCR8 monoclonal antibody and application thereof

A technology for monoclonal antibodies and applications, applied in the field of biomedicine or biopharmaceuticals, can solve the problems of lack of CCR8 monoclonal antibodies, small molecular weight, stability, immunogenicity and weak penetrability, etc.

Inactive Publication Date: 2020-02-25
普米斯生物技术(苏州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to the antigen reactivity of monoclonal antibodies, nanobodies also have some unique functional characteristics, such as small molecular weight, strong stability, good solubility, easy expression, weak immunogenicity, strong penetrability, and strong targeting , Humanization is simple, and

Method used

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  • Anti-CCR8 monoclonal antibody and application thereof
  • Anti-CCR8 monoclonal antibody and application thereof
  • Anti-CCR8 monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0153] Example 1: Expression and purification of human CCR8 protein

[0154] The nucleotide sequence of human CCR8 was synthesized in a commercially available vector, and the prepared plasmid was evenly mixed with the transfection reagent PEI 1:3, then allowed to stand for 20 minutes, and then added to appropriate cells (HEK293F cells), at 37°C, 6 %CO 2 Cultivate in a shaker incubator for 5-6 days; collect the cell supernatant and combine with Protein A beads at room temperature for 1 hour; wash the beads with phosphate buffer solution pH7.0, and then use 0.1M pH3.0Glycine to elute the protein;

[0155] Ultrafilter the eluted protein into PBS, measure the yield and take a sample for SDS-PAGE detection, and store the rest of the protein in a -80°C refrigerator;

Embodiment 2

[0156] Example 2: Preparation of CCR8 monoclonal antibody

[0157] In this example, a fully human antibody yeast screening platform was used to screen CCR8-specific monoclonal antibodies through CCR8+CHO cell stable transfection.

Embodiment 3

[0158] Preparation of Example 3 Nanobodies

[0159] A certain amount of hCCR8 antigen was mixed with an equal volume of Freund's adjuvant, and an alpaca was immunized once a week for several times in total to stimulate B cells to express antigen-specific nanobodies. After the immunization, 100mL peripheral blood lymphocytes were extracted and total RNA was extracted; cDNA was synthesized and VHH was amplified by nested PCR; and a phage display library was constructed.

[0160] Through screening and identification, multiple strains of nanobodies were obtained.

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Abstract

The invention relates to an anti-CCR8 monoclonal antibody and an application thereof. The invention discloses the anti-CCR8 monoclonal antibody. The antibody has a function of blocking binding betweenCCR8 and a ligand thereof, and the monoclonal antibody specifically targets Treg cells infiltrated in tumor tissues and having immunosuppressive activity, but does not target inflammatory the Treg cells and non-Treg type T cells. Compared with other targeting broad-spectrum Treg cell antibodies, the therapeutic antibody targeting CCR8 has better safety and effectiveness, so that the antibody is particularly suitable for immunotherapy of tumors.

Description

technical field [0001] The invention relates to the technical field of biomedicine or biopharmaceuticals, and more specifically relates to an anti-CCR8 monoclonal antibody and its application. Background technique [0002] With the successful marketing of multiple tumor immunomodulatory drugs targeting PD-1 / PD-L1, tumor immunotherapy has achieved revolutionary success. However, due to the infiltration of a large number of immunosuppressive cells in tumor tissue, the clinical response rate of PD-1 / PD-L1 tumor immunotherapy in patients with different types of tumors is still at a very low level. [0003] There are many reasons that limit the clinical response rate of PD-1 / PD-L1 immunotherapy, including the number of killer T cells infiltrating tumor tissue, the presence of a large number of immunosuppressive factors such as TGFβ, IDO, and adenosine in the tumor microenvironment, and the presence of Treg, Infiltration of various immunosuppressive cells such as MDSC in tumor ti...

Claims

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Application Information

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IPC IPC(8): C07K16/28A61K47/68A61K39/395A61K45/00A61P35/00
CPCA61K45/00A61K47/6849A61K2039/505A61P35/00C07K16/2866C07K2317/569C07K2317/732
Inventor 李斌伊刚
Owner 普米斯生物技术(苏州)有限公司
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