Purification method of F protein of human respiratory syncytial virus

Abstract

The invention discloses a purification method of F protein of human respiratory syncytial virus. The purification method comprises the following steps of: infecting HEp-2 cells or Vero cells with RSV (Respiratory Syncytial Virus), collecting the cells and a culture medium after complete pathological changes occur in the cells, centrifuging, taking supernatant, precipitating by using PEG6000, further purifying through sucrose density gradient centrifugation after the obtained precipitation is dissolved; subjecting a protein crude extracting solution to ion exchange chromatography and gel filtration chromatography in sequence for purifying the F protein after the envelope protein of the purified virus is dissolved in a lysis solution. The high-purified F protein can be obtained through two-step chromatography; the antigenicity of the F protein is kept in the purification process; and the purified F protein can be still detected by a group of monoclinic antibodies.

Description

technical field [0001] The invention belongs to the fields of virology and biotechnology, and in particular relates to a method for purifying F protein of human respiratory syncytial virus. Background technique [0002] Human respiratory syncytial virus (RSV) is the most important viral pathogen causing lower respiratory tract infections in infants and young children, especially bronchiolitis, worldwide. The peak period of onset in infants is from 6 weeks to 6 months after birth, especially within 2 to 6 months, and about 50% of infants are infected with RSV within one year of birth. The mortality rate for children with RSV infection is about 1%, and the mortality rate is higher for children with conditions such as stunting, chronic lung disease and congenital heart disease. In recent years, the epidemic incidence rate of RSV infection in my country is 0.24%~21.9%, and the hospital mortality rate is 0.5%~4%. Abnormal lung function, bronchial asthma and other lung diseases. ...

AI Technical Summary

Problems solved by technology

[0005] As a kind of membrane protein, F protein has many difficulties in its preparation, such as low content, low yield, loss of activity, and the need to optimize the purification process.

In terms of the purification of F protein, the current commonly used methods are purification methods such as immunoaffinity chromatography and protein electrophoresis, but these methods not only produce less protein but also cause partial loss of its antigenicity. The development of new vaccines urgently needs to establish a new method for the preparation and purification of F protein

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