Recombinant bifunctional fusion protein and application

A fusion protein, bifunctional technology, applied in the field of fusion proteins, can solve the problem of lack of bispecific antibodies

Pending Publication Date: 2021-04-13
南京华岩生物技术有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

To date there is no bispecific ant

Method used

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  • Recombinant bifunctional fusion protein and application
  • Recombinant bifunctional fusion protein and application
  • Recombinant bifunctional fusion protein and application

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preparation example Construction

[0034] The preparation method of the fusion protein is not particularly limited in the present invention, preferably including artificially synthesizing the nucleotide sequences encoding the polypeptide chains of each fusion protein, and cloning them into vectors respectively, so as to construct HY202-1 and HY202-2 respectively and HY202-3 expression vector; use the expression vector to electroporate CHO-K1 cells, induce protein expression and purify. In the present invention, there are no special limitations on the methods of electroporation, protein expression induction and purification, and conventional technical means in the field can be used.

[0035] The present invention also provides the application of the above-mentioned fusion protein in the preparation of medicine for treating MSLN-positive tumors. The fusion protein of the present invention can combine with MSLN and CD47 at the same time, thereby exerting a therapeutic effect on MSLN-positive tumor cells through an...

Embodiment 1

[0039] 1. Expression vector construction

[0040] Antibody heavy chain using Amatuximab monoclonal antibody VH MSLN -CH1-Fc (SEQ ID NO.10), the gene sequence was sent to Nanjing GenScript Synthetics (project number: C9962EE050-2); the V SIRPαD1 (SEQ ID NO.11) was fused with the anti-human MSLN antibody light chain (SEQ ID NO.12), and the gene sequence was sent to Nanjing GenScript (Project No.: C9962EE050-4) to obtain the polypeptide chain V SIRPαD1 -VL MSLN -CL (SEQ ID NO. 13). The synthesized genes were respectively cloned into vectors to constitute HY202-1 expression vectors.

[0041] Anti-human MSLN antibody heavy chain VH MSLN -CH1-Fc (same as above) and V SIRPαD1 For gene fusion, the gene sequence was sent to Nanjing GenScript for synthesis (project number: C9962EE050-6) to obtain the polypeptide chain VH MSLN -CH1-Fc-V SIRPαD1 (SEQ IDNO.14); Antibody light chain adopts Amatuximab monoclonal antibody VL MSLN -CL (SEQ ID NO.15), the gene sequence was sent to Nanji...

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Abstract

The invention provides a recombinant bifunctional fusion protein and application, and relates to the technical field of fusion proteins. The fusion protein is formed by connecting an anti-human MSLN antibody and a single-domain structure combined with human CD47 protein. The protein can be combined with MSLN and CD47 at the same time, so that the therapeutic effect on MSLN positive tumor cells is achieved through antibody-dependent, cell-mediated cytotoxicity and complement-dependent cytotoxicity activity, the combination of CD47 and SIRP on the surfaces of macrophages is blocked, the phagocytic effect of macrophages on tumor cells is activated, and the protein has remarkable anti-tumor activity. The tumor inhibition effect of the protein is better than that of single anti-MSLN monoclonal antibody treatment, and can be used for treating tumors which are invalid or tolerated by the anti-MSLN monoclonal antibody.

Description

technical field [0001] The invention belongs to the technical field of fusion proteins, in particular to a recombinant bifunctional fusion protein and its application. Background technique [0002] Mesothelin (mesotnelin, MSLN) is a 40kDa cell surface glycoprotein, not a cancer-specific antigen, but a differentiation antigen expressed at a relatively low level in normal tissues. Compared with normal tissues, mesothelin is highly expressed in many types of solid tumors, such as pancreatic cancer, ovarian cancer, malignant mesothelioma, lung adenocarcinoma, as well as endometrial, biliary, gastric and prostate cancers. Mesothelin is expressed on normal mesothelial cells lining the pleura, pericardium, and peritoneum. The limited distribution of mesothelin on normal tissues makes it a promising target for tumor-specific therapy. The mesothelin gene encodes a 71kDa precursor protein encoded by a 2138bp long cDNA containing 628 amino acids, which is processed into a 40kDa membr...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K39/395A61P35/00
CPCC07K16/30C07K14/70503A61P35/00C07K2317/31C07K2317/732C07K2317/76C07K2319/00A61K2039/505Y02A50/30
Inventor 姜东成王芳杜宜彬陈鹏刘志锋
Owner 南京华岩生物技术有限公司
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