A coxsackievirus A10 strain and a use thereof. Amino acid sequences of VP4, VP3, VP2 and VP1 proteins of the coxsackievirus A10 strain are respectively as represented by SEQ ID NOs 1, 2, 3 and 4, and amino acid sequences of 2A, 2B, 2C, 3A, 3B, 3C, and 3D proteins are respectively as represented by SEQ ID NOs 5, 6, 7, 8, 9, 10 and 11. The strain is obtained by using Vero cell isolation, readily infects Vero cells, and a higher titer can be obtained; the strain has advantages such as good immunogenicity, a strong cross neutralization capability, and genetic stability, and can be used for preparing a vaccine or drug for preventing or treating coxsackievirus A10 infection or a disease caused by coxsackievirus A10 infection.
A61P 31/14 - Antiviraux pour le traitement des virus ARN
G01N 33/569 - Tests immunologiquesTests faisant intervenir la formation de liaisons biospécifiquesMatériaux à cet effet pour micro-organismes, p. ex. protozoaires, bactéries, virus
A coxsackievirus A6 type strain, and an immunogenic composition and use thereof. The amino acid sequences of VP4, VP2, VP3 and VP1 proteins of the coxsackievirus A6 type strain are respectively shown in SEQ ID NO. 1, 2, 3 and 4. The amino acid sequences of 2A, 2B, 2C, 3A, 3B, 3C and 3D proteins are respectively shown in SEQ ID NO. 5, 6, 7, 8, 9, 10 and 11. The strain is obtained by separating Vero cells, is susceptible to Vero cells, can obtain higher titer, has the advantages of good immunogenicity, strong cross neutralization capacity etc., and can be used for preparing vaccines or drugs for preventing or treating a coxsackievirus A6 type infection or diseases caused by Coxsackievirus A6 infection.
A61K 39/39 - Préparations médicinales contenant des antigènes ou des anticorps caractérisées par les additifs immunostimulants, p. ex. par les adjuvants chimiques
A61P 31/14 - Antiviraux pour le traitement des virus ARN
3.
COXSACKIE VIRUS A16 STRAIN AND IMMUNOGENIC COMPOSITION AND APPLICATION THEREOF
Provided is a Coxsackie virus A16 strain containing a P1 structural protein and non-structural proteins 2A, 2B, 2C, 3A, 3B, 3C and 3D, the amino acid sequence of the P1 structural protein being as shown in SEQ ID NO. 1, and the amino acid sequences of the non-structural proteins 2A, 2B, 2C, 3A, 3B, 3C and 3D being as shown in SEQ ID NO. 4-10, respectively. The strain can be used for preparing a vaccine for preventing hand-foot-and-mouth disease caused by Coxsackie virus type A16.
Provided are a Hansenula engineered bacterium efficiently expressing a CA10 virus-like particle and use thereof. Said engineered bacterium comprise a recombinant vector carrying CA10 virus P1 and a 3CD genes. An original strain of the engineered bacterium is uracil-deficient Hansenula AU-0501. The P1 and 3CD genes are optimized according to a Hansenula preferred codon. Further provided is a method for preparing a CA10 virus-like particle, comprising: culturing an engineered bacterium, expressing the CA10 virus-like particle, and separating and purifying the virus-like particle by using ultrafiltration and a three-step chromatography process. Also provided is a hand-foot-and-mouth disease vaccine.
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.
C12N 15/81 - Vecteurs ou systèmes d'expression spécialement adaptés aux hôtes eucaryotes pour champignons pour levures
A61P 31/14 - Antiviraux pour le traitement des virus ARN
A61K 39/39 - Préparations médicinales contenant des antigènes ou des anticorps caractérisées par les additifs immunostimulants, p. ex. par les adjuvants chimiques
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 cell-density fermentation cultivation on recombinantly expressed engineered bacteria; inducing EV71 virus-like particle protein expression using methanol; collecting the bacteria by centrifugation and performing high-pressure homogenization for disruption; precipitating the supernatant with ammonium sulfate; and purifying by redissolution, ultrafiltration, ion exchange chromatography, size-exclusion chromatography, hydroxyapatite chromatography, etc.
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.
The present invention provides a method for preparing a genotype F mumps attenuated live vaccine. A mumps attenuated strain MHM-19 is inoculated to a human embryo lung diploid cell MHL-3 that is cultured to a monolayer, virus is released through the frozen-thawed cell in the peak of viral multiplication, cell debris is centrifuged and removed, a protective agent is added to a centrifuged virus suspension to obtain a semi-finished product of the vaccine, and the semi-finished product is frozen and dried to obtain the finished product of the genotype F mumps attenuated live vaccine. After the MHM-19 attenuated live vaccine prepared by the method of the present invention is injected into a rhesus monkey for immunizing, good humoral immunity is induced to be produced, and the detection results obtained at different time after immunization show that the MHM-19 attenuated live vaccine has immunizing potency superior to that of a reference mumps vaccine S79; in addition, the safety experiment shows that the rhesus monkey does not suffer from any parotitis-related symptom after being injected with the MHM-19 attenuated live vaccine, and the pathological tissue has no obvious difference from that of S79 strain by observing, so that the MHM-19 vaccine has high safety.
The present invention provides EV71 virus-like particles and a preparation method and application thereof. The method comprises: connecting a P1 protein gene and 3CD protease gene of an EV71 virus with PMV plasmid to construct PMV-P1-3CD recombinant expression plasmid; then putting the PMV-P1-3CD recombinant expression plasmid into a hansenula polymorpha AU-0501 expression bacterial strain to obtain an AU-PMV-P1-3CD recombinant expression bacterial strain; fermenting and culturing the recombinant expression bacterial strain and performing inducible expression on EV71 virus-like particle proteins by using methanol; centrifugally gathering thallus to perform homogenate crushing at high pressure; and purifying the supernate through ion-exchange column chromatography, hydrophobic chromatography, and sieve chromatography to obtain EV71 virus-like particles.
Provided is a method for determining free polysaccharide content in meningococcal Groups A, C, W135, and Y polysaccharide conjugate, comprising the following steps: 1) sampling: respectively taking 1 ml of a solution of meningococcal polysaccharide conjugate to be tested and a solution of a derivative corresponding to the meningococcal polysaccharide conjugate to be tested as Samples 1 and 3; and 2) centrifugation: additionally adding the solution of the meningococcal polysaccharide conjugate to be tested and the solution of the derivative corresponding to the meningococcal polysaccharide conjugate to be tested to a centrifugal tube; and adding a sodium deoxycholate solution to each centrifugal tube, standing for 30 min in an ice bath, then adding a hydrochloric acid solution, fully shaking, centrifuging, and collecting a supernatant from each centrifugal tube, as Samples 2 and 4; 3) calculating the polysaccharide concentration; and 4) calculating the free polysaccharide content. Through the present invention, the free polysaccharide content in meningococcal Groups A, C, W135, and Y polysaccharide conjugate can be accurately and conveniently determined, so as to evaluate the quality of a product.
Disclosed are a novel hepatitis A vaccine virus strain SH, a method for isolating the same, and a method for adapting the same to MRC-5 cells. The virus was isolated from the feces of a patient with an acute infection of hepatitis A and then transmitted to human diploid MRC-5 cells for adapted culture. In the adaptation process, the culture period for the early generation subculture was 35 days, and the culture period after the eighth passage was shortened to 24 days. With eight further passages, the antigen titer reached 1:512-1:1024, and the virus infectious titer reached 7.0-8.0lgCCID50/ml. Immunogenicity and cross protection assays showed that the hepatitis A inactivated vaccine produced from the virus strain has a good protective effect of immunogenicity. The virus strain is an ideal strain for the large-scale industrial production of hepatitis A inactivated vaccine.
A61P 31/14 - Antiviraux pour le traitement des virus ARN
A61P 1/16 - Médicaments pour le traitement des troubles du tractus alimentaire ou de l'appareil digestif des troubles de la vésicule biliaire ou du foie, p. ex. protecteurs hépatiques, cholagogues, cholélitholytiques
