Viral hepatitis treatment drug interferon production technology

Abstract

The invention discloses a viral hepatitis treatment drug interferon production technology which comprises the following steps: (1) acquisition of a target gene; (2) amplifying by PCR (polymerase chain reaction), to be more specific, in a microcentrifuge tube, using the 1 g / mL target gene obtained by the step (1) as template DNA, adding a right amount of a buffer solution, 4 kinds of dNTP (diethyl-nitrophenyl thiophosphate) mixtures, 0.5 2.5U / 50 muL DNA polymerase system, a pair of 0.1-0.5 mumol / L synthesized DNA primer to make the system pH 6.8-7.8; (3) gene recombination; (4) transformation; (5) separation and purification; (6) gene expression; (7) modification; and (8) freeze-drying, mass viral hepatitis treatment drug interferon can be produced in a short period of time, after modifying with polyethylene glycol, the half-life of the interferon is prolonged, hepatitis virus targeting property is improved, the antigenicity is weak, no human body immune response is caused, drug efficacy is good, production is fixed, stability is good, and the viral hepatitis treatment drug interferon production technology is suitable for popularization and application.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a production process of interferon, a medicine for treating viral hepatitis. Background technique [0002] The onset of viral hepatitis infection is mostly insidious, persistent infection can lead to liver cirrhosis and primary liver cancer, and the mortality rate is very high. For patients who eventually develop liver cirrhosis, the only effective treatment is liver transplantation, but in liver transplantation In the case that surgery cannot be popularized, early treatment of chronic viral hepatitis is particularly important. At present, there is no specific drug for the treatment of viral hepatitis, and interferon, nucleoside antiviral drugs and various Chinese patent medicines are mainly used, namely Immunoenhancing drugs, but the half-life of interferon on the current market is short, and the targeting is insufficient. What is more serious is that the natural interferon drug has a...

AI Technical Summary

Problems solved by technology

[0002] The onset of viral hepatitis infection is mostly insidious, persistent infection can lead to liver cirrhosis and primary liver cancer, and the mortality rate is very high. For patients who eventually develop liver cirrhosis, the only effective treatment is liver transplantation, but in liver transplantation In the case that surgery cannot be popularized, early treatment of chronic viral hepatitis is particularly important. At present, there is no specific drug for the treatment of viral hepatitis, and interferon, nucleoside antiviral drugs and various Chinese patent medicines are mainly used, namely Immunoenhancing drugs, but the half-life of interferon on the current market is short, and the targeting is insufficient. What is more serious is that the natural interferon drug has antigenic activity. The application number is CN200510069269.6, which discloses a hepatitis B genetic engineering targeting interferon And its production process, hepatitis B genetic engineering targeting interferon adopts the construction of human anti-HBsAg dsFv gene, which is connected with α-interferon (IFN-a) gene, and uses genetic engineering technology to express hepatitis B targeting interferon (dsFv- IFV fusion protein), the production process of hepatitis B genetic engineering targeting interferon, inoculate the prepared culture medium with the strain and then send it to the incubator for cultivation. After the completion of the cultivation, the bacteria multiply in a large number in the breeding room, and then the bacteria are collected by separation , and cleaved to separate the inclusion body protein, renatured in the folding solution, then purified, concentrated, and finally subpackaged and freeze-dried. After purification by affinity chromatography, it was proved that the fusion protein has interferon activity and reactivity with the surface antigen of hepatitis B virus , the fusion protein is of human origin, avoiding the harm caused by heterologousness, but the interferon is still antigenic, and the targeting is not enough