EV71 virus-like particles and hand-foot-and-mouth disease vaccine prepared from EV71 virus-like particles

Abstract

The invention relates to EV71 virus-like particles and hand-foot-and-mouth disease vaccine prepared from the EV71 virus-like particles. The EV71 virus-like particles are prepared according to the following steps: (1) constructing recombinant plasmid: respectively connecting pGAPZ alpha A plasmid with P1 protein gene of intestinal EV71 virus and 3CD protease gene to construct P1-pGAPZ alpha A and 3CD-pGAPZ alpha A recombinant plasmid; (2) transferring the recombinant plasmid to expression bacterial strains: sequentially transferring the recombinant plasmid to Pichia pastoris SMD1168 expression bacterial strains to obtain P1-pGAPZ alpha A-3CD-pGAPZ alpha A-SMD1168 recombinant expression strains; and (3) culturing thallus and purifying the EV71 virus-like particles: culturing the Pichia pastoris recombinant expression bacterial strains, centrifugalizing to separate supernate, and carrying out sucrose density gradient centrifugation on precipitation of the supernate after ultracentrifugation, thus obtaining the EV71 virus-like particles. In the invention, the EV71 virus-like particles are easy to obtain through thallus culture, have good stability, are easy to purify and suitable for preparing vaccine, and are convenient for industrial production.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an EV71 virus-like particle and a hand, foot and mouth disease vaccine prepared therefrom. Background technique [0002] Since 1957, hand, foot and mouth disease has broken out in many countries in the world, especially since the 1990s, it has become increasingly rampant in the Asia-Pacific region. Since 2007, there has been an outbreak of hand, foot and mouth disease in mainland China. By the end of September 2010, more than 3 million children have been reported to be infected with hand, foot and mouth disease, which seriously threatens the health and life safety of children. The pathogens of HFMD are mainly picornaviruses, members of the Enterovirus genus, mainly Coxsakie A group 16 (Coxsakie A 16 , CoxA16) and enterovirus 71 (enterovirus 71, EV71), wherein EV71 virus is the main pathogen that causes neurological complications and death cases. [0003] At present, there i...

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Problems solved by technology

However, since insect cells are mainly used to prepare VLPs at present, the culture conditions are relatively high; and because of intracellular expression, the VLP purification process is complicated, which limits the large-scale production requirements; and another major defect of the baculovirus-insect cell system It is the production of baculovirus particles and other pollutants that obviously affect the vaccine effect. Since the baculovirus particles are difficult to separate from the prepared VLP, the prepared VLP needs to be chemically inactivated and other measures used in clinical practice. These treatments may cause Affects the quality of the prepared VLP

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