Preparation method and application of ferroprotein nanostructure-based novel HER2 targeted therapeutic antibody

A technology of targeted therapy and nanostructure, applied in botany equipment and methods, biochemical equipment and methods, applications, etc., can solve problems such as high safety risks, achieve the effect of reducing safety risks and increasing protein loading

Inactive Publication Date: 2016-09-21
刘庆友 +6
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, injectable targeted therapeutic antibodies for

Method used

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Embodiment Construction

[0020] The preparation method of the novel HER2 (or ErbB2) targeting therapeutic antibody based on ferritin nanostructure of the present invention comprises the following steps in sequence:

[0021] 1) Construct a lentiviral expression vector of chicken ovalbumin OVA promoter nano-ferritin cage Ferritin, and package and produce toxin in 293T cells by liposome transfection, and concentrate and titer the virus liquid;

[0022] 2), then inject the virus liquid into the fresh fertilized embryos into the subgerminal cavity, and continue to hatch until the chickens are sexually mature; carry out genomic PCR identification on the seminal plasma and ovules of unfertilized eggs, and screen out positive chickens;

[0023] Or inject the virus liquid into the roosters at the end of the brooding period or the breeding period. After the roosters are fed for one month, take the chicken essence and observe it under a fluorescent microscope. The eggs were hatched, and some of the hatched chick...

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Abstract

The invention relates to a preparation method of a ferroprotein nanostructure-based novel HER2 targeted therapeutic antibody. The preparation method comprises the following steps: constructing lentiviral expression vectors of chicken egg albumin OVA promoter nano-Ferritin cages, transfecting the lentiviral expression vectors in 293 T-cells through lipidosome for packaging and producing toxin, and concentrating and titering virus liquid; then, injecting the virus liquid into subgerminal cavities of fresh fertilization embryos, and continuing hatching until chickens are hatched; after the chickens reach sexual maturity, performing genome PCR (Polymerase Chain Reaction) authentication on seminal plasma and ovules of unfertilized eggs, and screening positive chickens; mating G0-generation positive chickens and negative chickens of the opposite sex, performing the genome PCR authentication on the seminal plasma and ovules of the unfertilized eggs, after chickens reach sexual maturity, screening positive chickens, mating the positive chickens with negative chickens of the opposite sex, screening G2-geneartion positive chickens with the same method, and performing Western Blot and ELISA authentication on positive hens of each generation, so that a new variety of chickens which can be inherited stably and can stably express nano ferroprotein and monoclonal antibody fusion protein which can treat malignant tumors in egg white can be obtained. The preparation method has the advantages that the prepared protein is large in carrying capacity and can be taken orally.

Description

technical field [0001] The invention relates to a preparation method of a targeted therapeutic antibody, in particular to a preparation method and application of a novel HER2-targeted therapeutic antibody based on a ferritin nanostructure. Background technique [0002] Ferritin-based nanoparticles have the biochemical characteristics of high temperature resistance, non-denaturation, self-assembly and large specific surface area. Currently, injectable targeted therapeutic antibodies for the treatment of malignant tumors have high safety risks. Contents of the invention [0003] The purpose of the present invention is to overcome the above-mentioned defects of the prior art, and to provide a method for preparing a novel HER2-targeted therapeutic antibody based on a ferritin nanostructure that has a large protein load and can be taken orally. The purpose of the present invention is also to provide the application of the method. [0004] In order to achieve the above object,...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N15/13C12N5/10C07K16/32A61K39/395A61P35/00
CPCC07K16/32C07K2317/11C07K2317/24
Inventor 刘庆友李力石德顺张海航李志鹏陈忠毅温凯婷
Owner 刘庆友
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