A kind of humanized anti-aeg-1 single chain antibody and its application

A technology of AEG-1 and single-chain antibody, which is applied in the field of biomedicine and can solve problems such as immune damage

CN103408666BActive Publication Date: 2014-10-29FOURTH MILITARY MEDICAL UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2014-10-29

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Abstract

The invention discloses a humanized anti-AEG-1 single chain antibody and an application thereof. The single chain antibody comprises a heavy chain and a light chain, wherein the amino acid sequence of a variable region of the heavy chain is SEQ ID NO.1, and the amino acid sequence of a variable region of the light chain is SEQ ID NO.2, can specifically recognize AEG-1, has higher affinity and stronger immunocompetence, and has excellent targeting for cancer cells. The genes and amino acids of variable regions of the heavy chain and the light chain are verified through sequential analysis; the sequence of the single chain antibody has uniqueness. The single chain antibody not only provides support for anti-AEG-1 humanized genetically engineering antibody, but also establishes material base for diagnosis, targeted therapy, and prognosis of cancer.
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Description

technical field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to a humanized anti-AEG-1 single-chain antibody and its application. Background technique

[0002] Astrocyte elevated gene-1 (AEG-1), the encoded protein AEG-1, also known as metadherin (MTDH), is an oncogene discovered in recent years. Cancer, esophageal cancer, liver cancer, prostate cancer, lung cancer and other malignant tumor tissues are overexpressed, and are closely related to important biological characteristics such as tumor development, invasion and metastasis, and chemotherapy resistance.

[0003] Single chain Fv (single chain Fv, scFv) is a new type of genetically engineered antibody, which consists of the heavy chain variable region (V H ) and the light chain variable region (V L ) is connected by a short peptide (linker), and its advantages are: (1) It is an active antibody fragment with the smallest molecular weight, which is only 1 / 6 of the molecular...

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Embodiment Construction

[0038] The present invention will be further described in detail below in conjunction with specific embodiments, which are explanations of the present invention rather than limitations.

[0039] The present invention prepares and screens anti-AEG-1 single-chain antibodies based on ribosome display antibody library technology. The main operations include: extracting RNA from peripheral blood lymphocytes of lung cancer patients, and amplifying the V of the antibody by RT-PCR. H and V L Encoding gene, and then use a connecting peptide to connect the two into scFv gene. Linker peptides usually consist of 15 amino acids (Gly 4 Ser) 3 , to ensure that the peptide is flexible enough to facilitate V H and V L able to fold correctly.

[0040] The main process of preparing scFv is: first construct scFv (V H -Linker-V L ) DNA library; establish its ribosome display template; the template is transcribed and translated in an in vitro transcription and translation system to form a sc...