Method for preparing lactobionic acid-crosslinked hepatic cell targeted viral vector

A viral vector and hepatocyte technology, applied in the field of viral proteins, can solve problems such as poor specificity and unsatisfactory application, and achieve the effect of overcoming poor specificity

Inactive Publication Date: 2013-01-30
苏州百拓生物技术服务有限公司
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Problems solved by technology

Among them, viral vectors have high efficiency, but poor specificity, and the application has not been ideal

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  • Method for preparing lactobionic acid-crosslinked hepatic cell targeted viral vector
  • Method for preparing lactobionic acid-crosslinked hepatic cell targeted viral vector
  • Method for preparing lactobionic acid-crosslinked hepatic cell targeted viral vector

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Embodiment Construction

[0028] Taking rotavirus structural protein VP6 as a specific example below, the technical solution of the present invention is further described. The experimental methods described in the examples are conventional methods unless otherwise specified; the reagents and materials, if not specified, Both are commercially available. The same scheme is also applicable to rotavirus structural protein VP2, and various specific examples can be formed by selecting and replacing the corresponding conditions of each step, all of which are within the scope of protection of the present invention.

[0029] A method for preparing a lactosylated liver cell-targeting virus vector in this embodiment, such as figure 1 shown, including the following steps:

[0030] Step 1: Construct the gene of rotavirus (RV) structural protein VP6 into vector pET28a to obtain plasmid pET-VP6;

[0031] Step 2: Transform the plasmid pET-VP6 obtained in Step 1 into Escherichia coli to express the inclusion body pro...

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Abstract

The invention provides a method for preparing a lactobionic acid-crosslinked hepatic cell targeted viral vector. The method comprises the following steps of: I, constructing a gene of a rotavirus structural protein VP2 or VP6 onto a protein expression plasmid pET series vector to obtain an expression plasmid pET-VP2 or pET-VP6; II, transforming the expression plasmid pET-VP2 or pET-VP6 into escherichia coli, and giving expression to an inclusion body protein IB; III, performing renaturation and refolding on the inclusion body protein IB to form a protein R-VP2 or R-VP6; IV, crosslinking the protein R-VP2 or R-VP6 and an anti-cancer drug to form a conjugate; V, performing self-assembling on the conjugate to form a virus-like particle carrying the anti-cancer drug; and VI, covalently crosslinking targeted ligand lactobionic acid onto the outer surface of the virus-like particle carrying the anti-cancer drug to obtain the lactobionic acid-crosslinked hepatic cell targeted viral vector. According to the lactobionic acid-crosslinked hepatic cell targeted viral vector prepared by the method provided by the invention, a hepatocellular carcinoma cell HepG2 can be specifically identified, so that the carried drug can be released in the hepatocellular carcinoma cell and generates cytotoxicity, and a new way is provided for targeted drug treatment of hepatocellular carcinoma.

Description

technical field [0001] The invention belongs to the technical field of viral proteins, and in particular relates to a preparation method of a lactosylated liver cell-targeting viral vector. Background technique [0002] Liver cancer is one of the most common tumors, and its incidence is increasing worldwide, seriously endangering human life and health. The current methods of treating liver cancer mainly include chemotherapy, surgery, intervention and other means, but the effect is not satisfactory. With the development of genetic science and technology, gene therapy for liver cancer has become possible. The key issue of gene therapy is the carrier of the gene. At present, the technology in this field is developing rapidly. The commonly used gene therapy carriers for liver cancer mainly include viral vectors, liposomes, plasmids, and cationic polymers. Among them, viral vectors have high efficiency, but poor specificity, and their application has not been ideal. [0003] L...

Claims

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Application Information

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IPC IPC(8): A61K47/48A61K47/26A61K45/00A61P35/00A61K47/64
Inventor 赵庆欢刘红海乐晓桐李咏梅顾莉娟
Owner 苏州百拓生物技术服务有限公司
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