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Light-chain variable region and heavy-chain variable region of FMU-EPCAM-4F6 monoclonal antibody

A technology of FMU-EPCAM-4F6, 1. FMU-EPCAM-4F6, applied in the field of monoclonal antibodies, can solve the problems of antibody drug hypersensitivity and the like

Active Publication Date: 2013-02-13
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Claims
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Problems solved by technology

However, when the mouse-derived antibody drug is used in the human body, due to its immunogenicity, it will cause an immune response in the human body, resulting in the clearance of the antibody drug or hypersensitivity mediated by immune complexes

Method used

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  • Light-chain variable region and heavy-chain variable region of FMU-EPCAM-4F6 monoclonal antibody
  • Light-chain variable region and heavy-chain variable region of FMU-EPCAM-4F6 monoclonal antibody
  • Light-chain variable region and heavy-chain variable region of FMU-EPCAM-4F6 monoclonal antibody

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Embodiment Construction

[0030] The applicant immunized BALB / c mice with recombinant human EPCAM, prepared a group of mouse anti-human EPCAM monoclonal antibodies, and screened the FMU-EPCAM-4F6 monoclonal antibody hybridoma cell line that can stably secrete high-affinity anti-human EPCAM, Ascites fluid was prepared to obtain high-affinity anti-human EPCAM monoclonal antibody. Confirm the uniqueness of the gene sequence and corresponding protein sequence and its CDR sequence; provide support for anti-human EPCAM chimeric or humanized genetic engineering antibodies.

[0031] The present invention will be described in detail below in conjunction with the accompanying drawings, which are explanations of the present invention rather than limitations.

[0032] The present invention is specifically implemented according to the following steps:

[0033] 1 Preparation of mouse anti-human EPCAM high-affinity antibody

[0034] 1.1 Preparation and purification of monoclonal antibodies

[0035] Design primers ...

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Abstract

The invention discloses a light-chain variable region and a heavy-chain variable region of an FMU-EPCAM-4F6 monoclonal antibody, wherein an amino acid sequence of the light-chain variable region of the FMU-EPCAM-4F6 monoclonal antibody is shown as SEQ ID NO.1, and an amino acid sequence of the heavy-chain variable region of the FMU-EPCAM-4F6 monoclonal antibody is shown as SEQ ID NO.2. In the invention, a group of mouse anti-human EPCAM monoclonal antibodies is prepared by a recombinant human EPCAM immune BALB / c mouse, hybridoma cell strains which can stably secrete high-affinity anti-human EPCAM monoclonal antibodies are screened, and the high-affinity anti-human EPCAM monoclonal antibodies are obtained by preparing ascitic fluid. The invention confirms the uniqueness of a gene sequence and a corresponding protein sequence as well as a CDR sequence and provides support for anti-body EPCAM chimeric or humanized gene engineering antibodies.

Description

technical field [0001] The present invention belongs to the technical field of biomedicine and relates to a monoclonal antibody, in particular to a light chain and heavy chain variable region of an anti-human EPCAM FMU-EPCAM-4F6 monoclonal antibody, including its amino acid sequence and nucleotide sequence . Background technique [0002] Epithelial cell adhesion molecule (EPCAM) was originally discovered as a dominant epitope in colon cancer, and it was considered to be a reliable binding site for cell adhesion and therapeutic antibodies. Current studies believe that EPCAM is a glycosylated type I transmembrane glycoprotein with a molecular weight of 30 to 40kD. Its structure includes an epidermal growth factor-like and a thyroglobulin-like extracellular region, a transmembrane region and a Cytoplasmic domain of 26 amino acids. In normal cells, EPCAM is mainly located in the tight junction of the epithelial cell space. As shown by the intensity and frequency of EPCAM expr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/18C12N15/13G01N33/577
Inventor 金伯泉杨琨张葵宋朝君孙元杰朱参胜谢鑫
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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