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Monoclonal antibody against TSPAN8 and application of monoclonal antibody against TSPAN8

An antibody and variable region technology, applied in the field of biomedicine, can solve the problems of lack of fully human antibodies, etc., and achieve the effects of strong specificity, growth inhibition, and high affinity

Active Publication Date: 2020-01-31
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] At present, there is still a lack of fully human antibodies against TSPAN8 with high affinity anti-tumor effects in China

Method used

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  • Monoclonal antibody against TSPAN8 and application of monoclonal antibody against TSPAN8
  • Monoclonal antibody against TSPAN8 and application of monoclonal antibody against TSPAN8
  • Monoclonal antibody against TSPAN8 and application of monoclonal antibody against TSPAN8

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Embodiment

[0052] A method for obtaining a monoclonal antibody against TSPAN8. In this embodiment, the constant region of the heavy chain of the IgG1 subtype and the constant region of the light chain comprising kappa are selected. The brief description includes the following steps:

[0053] S1: The gene sequence encoding the extracellular segment of TSPAN8 (TSPAN8-LEL) and the C-terminal FC-tag were cloned into the pFUSE expression vector by gene recombination technology, and the human TSPAN8-LEL-FC fragment was constructed and expressed in HEK293F cells Obtain TSPAN8-LEL-FC fusion protein;

[0054] S2: Using the TSPAN8-LEL-FC fusion protein as the antigen, through three rounds of antibody library enrichment screening, eight high-affinity ScFv sequences were obtained from the fully human ScFv phage library, which were named TsC01-TsC08.

[0055] S3: The above eight positive ScFv sequences, namely TsC01-TsC08, were cloned into eukaryotic expression vectors, expressed and purified after t...

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Abstract

The invention discloses a monoclonal antibody against TSPAN8 and application of the monoclonal antibody against TSPAN8. The antibody can be specifically bound to TSPAN8 protein; a heavy-chain variableregion of the antibody is encoded by a nucleotide sequence as shown in SEQ ID NO:1 or a homologous sequence of the nucleotide sequence as shown in SEQ ID NO:1; and a light-chain variable region of the antibody is encoded by a nucleotide sequence as shown in SEQ ID NO:2 or a homologous sequence of the nucleotide sequence as shown in SEQ ID NO:2, and the antibody can be used for producing a medicine composition for treating and / or preventing a TSPAN8-mediated disease capable of promoting a molecular regulation mechansim of epithelial-mesenchymal transition and stemness enhancement of tumor cells and a key molecular target. The provided monoclonal antibody against TSPAN8 has high affinity, is high in specificity and good in stability, has the capability of inhibiting proliferation and migration of a breast cancer cell line, and can inhibit growth of tumors.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a fully human anti-TSPAN8 monoclonal antibody and its application. Background technique [0002] Tetraspanins, also known as the tetraspanins superfamily (TM4SF), consist of 33 family members, including TSPAN8, CD63, CD37, CD53, CD81 and CD9 related to differentiation (CD). The common feature of the Tetraspanins family is that they have four highly conserved membrane spanning domains, which play important biological regulation in various cellular functions. [0003] Tumor invasion, metastasis and drug resistance are the main causes of clinical treatment failure and patient death. In the tumor microenvironment (Tumor Microenvironment, TME), the tetraspanning protein TSPAN8 promotes the molecular regulation mechanism and key molecular targets of tumor cell epithelial-mesenchymal transition (Epithelial Mesenchymal Transition, EMT) and stemness (Stemness). Accumulating evidence shows tha...

Claims

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Application Information

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IPC IPC(8): C07K16/28C12N15/13A61K39/395A61P35/00A61P35/04
CPCA61K2039/505A61P35/00A61P35/04C07K16/28C07K2317/73C07K2317/92
Inventor 王红霞葛伟玉杨光朱容萱马培祥周天浩李军建许静轩
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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