Collagen-specific single-chain antibody, its coding gene and application

A specific binding, single-chain antibody technology, applied in the direction of antibodies, hybrid peptides, specific peptides, etc., can solve the problems of reducing the therapeutic effect of full-length antibodies, slow speed, insurmountable and other problems, and achieve the effect of good application prospects.

Active Publication Date: 2019-01-18
DUBU BIOSCIENCES
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, due to the large molecular weight of this kind of full-length antibody, it is difficult to overcome the internal pressure of the tumor, and the speed of penetrating blood vessels to reach the tumor site is relatively slow. Obviously; these factors greatly reduce the therapeutic effect of full-length antibodies

Method used

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  • Collagen-specific single-chain antibody, its coding gene and application
  • Collagen-specific single-chain antibody, its coding gene and application
  • Collagen-specific single-chain antibody, its coding gene and application

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Embodiment Construction

[0032] As mentioned above, in view of the many defects in the prior art, the inventors of this case have prepared a single-chain antibody (CBD-scFv) with collagen-specific binding through long-term research and a lot of practice. For example, the functional region of cetuximab is fused and expressed, and the yeast expression system is preferably used to successfully obtain the CBD-scFv fusion antibody fragment. The CBD-scFv has good collagen-specific binding ability in vitro and in vivo, and Maintaining the ability to bind to EGFR and anti-tumor activity, CBD-scFv can quickly accumulate at the site of tumor occurrence and play a killing role on tumor cells.

[0033] The technical solutions of the present invention will be described in more detail below in conjunction with the accompanying drawings and preferred embodiments.

[0034] 1. High-efficiency expression and purification of CBD-scFv in yeast

[0035] i, construction of CBD-scFv expression vector

[0036] V of CBD-scF...

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Abstract

The invention discloses a single-chain antibody specifically bound with collagen and coding genes thereof and application. In a better embodiment, the single-chain antibody (CBD-scFv) is mainly formed by integrating a polypeptide CBD specifically bound with the collagen with an scFv functional domain of cetuximab. Further more, the single-chain antibody specifically bound with the collagen is formed by coding the genes with the sequences shown in SEQ ID NO:1. According to the single-chain antibody specifically bound with the collagen and the coding genes thereof and the application, the collagen binding domain CBD polypeptide and the functional domain scFv of the cetuximab are integrated through a gene engineering method, and a yeast expression system is preferably utilized; in-vitro experiments and in-vivo experiments prove that the CBD-scFv can be specifically bound with a collagen institution, the binding capacity of the cetuximab with EGFR on the surface of a tumor cell is kept, the very good killing effect on tumors in vitro is achieved, the CBD-scFv can gather on tumor occurrence portions enriched with the collagen in a targeted mode in vivo and can be targeted to the tumor portions more quickly relative to a full-length antibody, and the single-chain antibody has the good application prospect in the fields of tumor targeting therapy and medicine carrying.

Description

technical field [0001] The invention relates to a therapeutic antibody, in particular to a single-chain antibody with collagen-specific binding ability, its preparation method and its application in tumor treatment. Background technique [0002] Collagen is the main component of the extracellular matrix, and about 30% of it is collagen in animals. Collagen plays an important role in the process of cell adhesion, migration, differentiation, and proliferation. A large number of studies have shown that in tumor cells Collagen is overexpressed in the place where it occurs relative to normal tissue, and collagen provides signal transduction and physical scaffolding for tumor cells during the occurrence and metastasis of tumors. At present, more and more drugs targeting tumor cell surface targets have been discovered and studied, and good tumor cell killing effects have been obtained in vitro experiments, but in vivo experiments due to the formation of extracellular matrix overexp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62G01N33/68G01N33/577G01N33/569A61K39/395A61P35/00
Inventor 戴建武梁辉陈冰
Owner DUBU BIOSCIENCES
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