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Anti-PD1-CD19 bispecific antibody, pharmaceutical composition thereof and application thereof

A PD1-CD19, bispecific antibody technology, applied in the field of its pharmaceutical composition, anti-PD1-CD19 bispecific antibody, can solve the problem of weakening the tumor immune effect, achieve enhanced anti-tumor effect, small molecular weight, enhanced non-specific The effect of heterosexual killing

Pending Publication Date: 2020-11-17
SHUNHAO CELL BIOTECHNOLOGY (TIANJIN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The monoclonal antibody binds to the effector cell surface active receptor PD-L1 through the Fc segment, thereby mediating the killing effect, but the T cells with immune killing effect cannot be effectively mediated because of the lack of the above-mentioned receptors on the surface, thus weakening the body's anti-tumor effect. immune effect

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0023] Construction of double antibody molecules

[0024] In the present invention, the anti-PD1-CD19 bispecific antibody is constructed by using the CD19 monoclonal antibody IgG and the scFv of the PD1 monoclonal antibody in series. The light chain variable region VL of the anti-PD1 monoclonal antibody and the heavy chain variable region VH are connected through a peptide linker L1 to obtain the anti-PD1 single-chain antibody fragment VL-L1-VH, namely the anti-PD1 fragment scFv1. L2 is used to link the single-chain antibody fragment with the heavy chain of the anti-CD19 monoclonal antibody to obtain a bispecific antibody molecule.

Embodiment 2

[0026] Expression and purification of double antibody

[0027] The heavy chain and light chain DNA fragments of the double antibody were subcloned into the pTT5 vector, and the recombinant plasmids were extracted and co-transfected into CHO cells and / or 293E cells. After the cells have been cultured for 5-7 days, the culture medium is subjected to high-speed centrifugation and vacuum filtration through a microporous membrane, then loaded onto a HiTrap MabSelectSuRe column, and the protein is eluted in one step with an eluent containing 100mM citric acid, pH 3.5, and the target is recovered Samples were dialyzed into PBS pH 7.4.

[0028] The purified protein was detected by HPLC. The purified antibodies were added to non-reducing electrophoresis buffer for detection by SDS-polyacrylamide gel electrophoresis; the purified double antibodies were respectively added to reducing electrophoresis buffer and boiled for detection by SDS-polyacrylamide gel electrophoresis.

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Abstract

The invention relates to the field of antibody drugs, and particularly relates to an anti-PD1-CD19 bispecific antibody, a pharmaceutical composition thereof and application thereof. The antibody comprises a heavy chain and a light chain, wherein the heavy chain and the light chain both comprise variable regions, and the variable regions comprise complementary determining regions. The antibody further comprises an immunoglobulin antibody IgG and a variable fragment scFv, wherein the scFv comprises a variable region VH and a variable region VL. According to the antibody, two specific single-chain antibodies are connected in series by using a molecular biological technology, one end of an anti-epidermal growth factor receptor is used for recognizing tumor cells, and the other end of the anti-epidermal growth factor receptor is used for recognizing and activating T lymphocytes by using an anti-PD1 single-chain antibody, so that the effect of removing the tumor cells is achieved.

Description

technical field [0001] The present invention relates to the field of antibody medicines, in particular to an anti-PD1-CD19 bispecific antibody, its pharmaceutical composition and its use. Background technique [0002] PD1 gene is a 55kDa transmembrane protein, its molecule is composed of 4 subunits: δ, ε, γ, ζ, the respective molecular masses are 18.9kDa, 23.1kDa, 20.5kDa, 18.7kDa, which is an Ig gene A member of the super family. PD1 is an inhibitory member of the CD28 family expressed on activated B cells, T cells and myeloid cells. [0003] CD19 is a 95 kDa membrane receptor that is expressed early in B cell differentiation and continues to be expressed until B cells are triggered for terminal differentiation. The extracellular domain of CD19 contains two C2-type immunoglobulin (IG)-like domains separated by a smaller potentially disulfide-linked domain. The CD19 cytoplasmic domain is structurally unique but highly conserved among humans, mice and guinea pigs (Fujimoto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/46A61K39/395A61P35/00
CPCA61P35/00C07K16/2803C07K16/2818C07K2317/31C07K2317/56C07K2317/622
Inventor 李锋赵晓翠胡亚丽
Owner SHUNHAO CELL BIOTECHNOLOGY (TIANJIN) CO LTD
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