Abstract: The present invention provides a Hansenula engineering fungus that efficiently expresses CA10 virus-like particles and use thereof. The engineering fungus comprises a recombinant vector carrying P1 and 3CD genes of the CA10 virus, and the starting strain of the engineering fungus is uracil auxotroph. The Hansenula AU-0501, P1 and 3CD genes are optimized according to preferred codons of Hansenula. The present invention also provides a preparation method of CA10 virus-like particles, comprising: culturing engineering fungi, expressing CA10 virus-like particles, and separating and purifying virus-like particles by ultrafiltration and three-step chromatography.

Abstract: The present invention provides a method for purifying an EV71 virus-like particle and a method for preparing a vaccine thereof. The virus-like particle is obtained by performing high density fermentation cultivation on recombinantly engineered bacteria; inducing expression of the EV71 virus-like particle protein expression using methanol; collecting the bacteria by centrifugation and performing high-pressure homogenization for disruption; performing precipitation on the supernatant with ammonium sulfate; and purifying by redissolution, ultrafiltration, ion exchange chromatography, molecular sieve chromatography, hydroxyapatite chromatography, etc.

Abstract: The present invention provides a method for purifying an EV71 virus-like particle and a method for preparing a vaccine thereof. The virus-like particle is obtained by performing high density fermentation cultivation on recombinantly engineered bacteria; inducing expression of the EV71 virus-like particle protein expression using methanol; collecting the bacteria by centrifugation and performing high-pressure homogenization for disruption; performing precipitation on the supernatant with ammonium sulfate; and purifying by redissolution, ultrafiltration, ion exchange chromatography, molecular sieve chromatography, hydroxyapatite chromatography, etc.

Abstract: The present invention provides EV71 virus-like particles and a preparation method and application thereof. The method comprises: connecting a P1 protein gene and a 3CD protease gene of an EV71 virus with a PMV plasmid to construct a PMV-P1-3CD recombinant expression plasmid; then transforming a Hansenula polymorpha AU-0501 expression strain with the PMV-P1-3CD recombinant expression plasmid to obtain an AU-PMV-P1-3CD recombinant expression strain; fermenting and culturing the recombinant expression strain, and inducing the recombinant expression strain to express the EV71 virus-like particle protein with methanol; centrifuging and collecting mycelia for homogeneous breakage at a high pressure; and purifying the supernatant through ion-exchange chromatography, hydrophobic chromatography, and molecular sieve chromatography, so as to obtain EV71 virus-like particles.

Abstract: The present invention provides EV71 virus-like particles and a preparation method and application thereof. The method comprises: connecting a P1 protein gene and a 3CD protease gene of an EV71 virus with a PMV plasmid to construct a PMV-P1-3CD recombinant expression plasmid; then transforming a Hansenula polymorpha AU-0501 expression strain with the PMV-P1-3CD recombinant expression plasmid to obtain an AU-PMV-P1-3CD recombinant expression strain; fermenting and culturing the recombinant expression strain, and inducing the recombinant expression strain to express the EV71 virus-like particle protein with methanol; centrifuging and collecting mycelia for homogeneous breakage at a high pressure; and purifying the supernatant through ion-exchange chromatography, hydrophobic chromatography, and molecular sieve chromatography, so as to obtain EV71 virus-like particles.