Single-chain antibody targeting EGFRvIII and application of antibody

A technology of single-chain antibody and light chain, which can be used in medical preparations of non-active ingredients, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-tumor drugs, etc., and can solve the loss of coding base pairs and other problems to achieve the effect of high specificity, low lethality, and reduced clearance rate

Active Publication Date: 2021-04-16
GUIZHOU MEDICAL UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

This mutation originates from the in-frame deletion of exons 2-7 in the extracellular region, resulting in the loss of 841 coding base pairs and a glycoprotein with a molecular weight of only 145 kDa. Due to the lack of extracellular L1 and CR1 (rich Cysteine) subdomain and therefore not capable of being activated by any ligand

Method used

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  • Single-chain antibody targeting EGFRvIII and application of antibody
  • Single-chain antibody targeting EGFRvIII and application of antibody
  • Single-chain antibody targeting EGFRvIII and application of antibody

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Experimental program
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Effect test

Embodiment

[0037] Screening and preparation of single-chain antibodies:

[0038] 1. Immunization of animals

[0039] First, 10 8-week-old BALB / c mice were immunized with EGFRvIII polypeptide (LEEKKGNYVVTDHC) and adjuvant, and divided into experimental group (EGFRvIII polypeptide and adjuvant mixed at 1:1, dosage of 100 μg / mouse) and control group (adjuvant dosage 100μg / only).

[0040] 2. ELISA detection of immune results

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Abstract

The invention relates to the technical field of genetic engineering antibodies, in particular to a single-chain antibody targeting EGFRvIII and application. According to the anti-EGFRvIII single-chain antibody prepared and screened by utilizing genetic engineering, the obtained anti-EGFRvIII single-chain antibody is relatively high in specificity, has good biological activity, and can be effectively combined with EGFRvIII protein. After the anti-EGFRvIII single-chain antibody disclosed by the invention is coupled with an anti-tumor drug, an antibody coupling drug has good targeting to various cancer cell tissues expressing EGFRvIII, has a good killing effect on target cells with EGFRvIII expression, and has relatively low killing effect on non-target cells; and compared with a complete antibody, the anti-EGFRvIII single-chain antibody and drug conjugate disclosed by the invention can reduce the in-vivo clearance rate of the complete antibody and drug conjugate and prolong the half-life period, so that the in-vivo utilization rate of the drug is improved, and the anti-EGFRvIII single-chain antibody and drug conjugate has better in-vivo pharmacokinetic properties.

Description

technical field [0001] The invention relates to the technical field of genetic engineering antibodies, in particular to a single-chain antibody targeting EGFRvIII and its application. Background technique [0002] EGFR is a transmembrane glycoprotein with a molecular weight of about 170kDa, also known as ErbB or HER receptor, and is a member of the epidermal growth factor receptor family. EGFR family members are involved in important signaling pathways of angiogenesis, cell differentiation, proliferation, and survival in many cancer processes, and in normal cell growth and malignant transformation, prevention of apoptosis, drug resistance, tumor stem cells, and metastasis of various cancers Play an important role. EGFR has many mutants, the most common of which is EGFRvIII. This mutation originates from the in-frame deletion of exons 2-7 in the extracellular region, resulting in the loss of 841 coding base pairs and a glycoprotein with a molecular weight of only 145 kDa. D...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28A61K47/68A61P35/00
Inventor 刘亭张煜彬官志忠王永林李勇军
Owner GUIZHOU MEDICAL UNIV
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