186 results about "Rabies vaccine" patented technology

The present invention relates to a method for purifying recombinant human serum albumin (rHSA) protein. The method comprises the following steps: fermented liquid containing rHSA is processed by a ceramic membrane, supernatant liquid is orderly purified by high salt cation exchange chromatography, hydrophobic layer exchange chromatography and weak anion exchange chromatography, and purified rHSA is obtained. The present invention is characterized in that solution processed by high salt cation exchange chromatography is processed by borate and then filtered by hollow fibers. The rHSA obtained can be used for producing vaccines for humans against viruses with a cell culture method, particularly rabies vaccines.

The invention provides a method for producing rabies viruses by the suspension culture of BHK21 cells, which comprises a step of performing suspension culture of BHK21 cells in bioreactor containing a BHK21 cell culture medium, wherein the conditions for the suspension culture in the step include a temperature of 32 to 36 DEG C, a pH value of 7.0 to 8.0 and a dissolved oxygen concentration of 30 to 50 percent; and the BHK21 cell culture medium comprises the components shown in a table 1. The method overcomes the biases of the prior art and realizes the production of the rabies viruses by the suspension culture of BHK21 cells in the bioreactor. The obtained rabies viruses can be used for producing rabies vaccine. Due to the automatic culture environment parameter control of the bioreactor, the cells grow and the viruses propagate in more favorable environments, the virus titer is improved and the large-scale automatic continuous production can be realized.

The invention provides a method for culturing baby hamster kidney (BHK) 21 cell in a serum-free way, which leads the BHK 21 cell to be inoculated into a cell culture medium for culturing, wherein the cell culture medium comprises a basic culture medium and further comprises 100-200g/100L of soy protein, 100-200g/100L of pea protein, 100-200g/100L of broad bean protein, 0-100g/100L of potato protein, 0-200g/100L of wheat gluten protein and 50-100g/100L of rice protein by taking the volume of solvent of the culture medium as reference. In addition, the invention also provides a vaccine (such asrabies vaccine and foot-and-mouth disease vaccine) preparation method comprising the method for culturing the BHK 21 cell. The BHK 21 cell cultured by the culture medium containing the vegetable protein is high in culture density, beneficial to separating down-stream products of the cells, low in cost, small in batch difference, good in safety and suitable for producing virus host, expression vector and the like of biological products such as vaccine and the like.

The invention relates to the field of biotechnology, in particular to a virus seed for producing vaccines for preventing human rabies by utilizing human diploid cells (KMB17) and a preparation method thereof. In the invention, a rabies fixed virus CTN-1V5 strain is continuously subcultured in the human diploid cells (KMB17), and a terminal dilution method is used for screening viruses with highertiter, thereby obtaining a rabies virus strain which is suitable for the human diploid cells (KMB17) and has good immunogenicity and heredity stability, and culturing a rabies vaccine virus seed (CTN-DK strain) capable of efficiently reproducing in the human diploid cells (KMB17). By using the virus seed for producing a human diploid cell (KMB17) rabies vaccine, the risk caused by residual heterogonous DNA (deoxyribonucleic acid) in the vaccine which is currently used at home can be effectively avoided, and the safety and practicability of the rabies vaccine in China are further improved, thus the invention has great social and economic benefits.

The invention relates to a freeze-dried rabies vaccine for humans and a preparation method of the vaccine, relates to the field of vaccine production preparation technologies and aims at solving the problems that effective virus antigen expression content is low, the side effect rate of a vaccinator is high and vaccine yield and quality can not meet standard requirements as only a biological reactor is adopted for producing a rabies vaccine. The freeze-dried rabies vaccine for humans is obtained by inoculating aG strain rabies virus on Vero cells and sequentially carrying out ultrafiltration and concentration, separation and purification as well as freeze drying, wherein the packing volume of the freezed-dried rabies vaccine for human use is 0.5ml/dose, and during freeze drying, the adopted vaccine freeze-drying protecting agent comprises the following ingredients: 60-90g/l of trehalos, 6-14g/l of sodium glutamate, 3-6g/l of urea, 2-3g/l of L-arginine and 10g/l of 199 culture medium, and the vaccine freeze-drying protecting agent does not contain gelatin, human serum albumin or dextran. The freeze-dried rabies vaccine for humans has the advantages that cost is low, operation is easy, pollution is hardly produced, vaccine quality and yield are greatly improved, the content of impurities in a vaccine is reduced, allergy reactions are hardly caused, and vaccine safety is greatly improved.

The invention relates to a novel rabies vaccine and a preparation method thereof. Based on a vaccine carrier for chimpanzee type adenovirus AdC68 genome, an inventor constructs a novel rabies vaccine carrier. The invention further provides a virus vaccine prepared from the vaccine carrier, which can efficiently express and has excellent immunogenicity.

The invention provides a rabies vaccine for human beings, which contains an outer membrane fragment of a split rabies virus particle and tetanus toxoid; the outer membrane fragment comprises furcella and matrix protein, wherein each dosage of rabies vaccine for human beings contains 10-150 micrograms of virus protein; and the dosage of the tetanus toxoid is 3-10 Lf/dosage. Compared with the existing virus purification stock solution, the rabies vaccine for human beings contains less other proteins, has higher immunogenicity and effect, and is more secure for use.

The invention discloses a human diploid cell rabies vaccine and a preparation method thereof, and belongs to the field of vaccines. A CDKHBP-1 strain is inoculated into a human diploid cell WI-38, and the rabies vaccine is obtained through separation and purification. The human diploid cell does not contain any contaminants or oncogenicity, and is obvious superior to an animal cell serving as a medium for producing the vaccine; and the method is suitable for large-scale industrial production, a purification process is perfect, and the vaccine contains a few impurities and has high purity and immunogenicity.

A purified diplontic cell vaccine in the form of freeze-dried powder injection or liquid injection for rabies is prepared through inoculating rabies virus into human diploied cells.

The present invention provides an adjuvant, which includes at least one single strand deoxynucleotide containing a CpG dinucleotide. The single strand deoxynucleotide comprises one or more CpG dinucleotides. When used in combination with rabies vaccine, HBV vaccine or other vaccines, the adjuvant can significantly improve the immune effect of the vaccine.

The invention discloses a human vaccine for preventing hydrophobia and tetanus, which belongs to the field of vaccines. The vaccine consists of a hydrophobia vaccine and a tetanus toxin, wherein the hydrophobia vaccine is obtained by inoculating a CDKHBP-1 strain onto a human diploid cell WI-38 and purifying. By using the hydrophobia vaccine and the tetanus toxin prepared with the preparation method disclosed by the invention together, immunity to hydrophobia and tetanus can be realized; and moreover, the tetanus toxin can be used for remarkably enhancing valence effect of the hydrophobia vaccine, plays a role in enhancing immunity, and is convenient for administration.

The invention provides rabies vaccine for human being. The vaccine comprises an outer membrane segment of cracked rabies complete viral particles and tetanus toxoid, and the outer membrane segment comprises spike and matrix protein. Each dose of the rabies vaccine comprises 10 to 100 micrograms of viral protein; the dosage of the tetanus toxoid is 2 Lf/dose to 10 Lf/dose; and the dosage of aluminum phosphate adjuvant is 0.5mg/dose. Compared with the existing virus purified stock solution, the rabies vaccine for human being has the advantages that the content of impurity protein is greatly reduced, the immunogenicity and the potency are high, and the rabies vaccine is safer to use.

The invention belongs to the technical field of biological engineering, in particular to a purification method during vaccine production, solving the problems that DNA is difficult to be removed and protein is easy to be polymerized and precipitated because column chromatography method is singly adopted during the previous production process of the hydrophobia vaccine. The method comprises the following steps: adopting a 750KD hollow fibre ultrafiltration column or a 300KD ultrafiltration membrane for concentrating virus harvested liquid or removing part of impurities, adopting non-restriction endonuclease for degrading DNA, and then removing nuclease by the method of ultrafiltration or column chromatography. The method is characterized by easy magnification, high product purity and obvious DNA removing effect, and greatly improves the safety of the vaccine.

The invention aims at providing a rabies virus resistant neutralizing antibody, and particularly provides a humanized or completely humanized monoclonal antibody to meet the requirement for clinically diagnosing and/or treating rabies. A phage antibody library is prepared by adopting a phage antibody library technology and taking 32 parts of high-potency healthy human peripheral blood inoculated with rabies vaccines as a raw material; 7 ELISA positive antibodies are obtained through three rounds of screening from the phage antibody library; and furthermore, the neutralizing activity of the 7 ELISA positive antibodies is measured through an RFFIT method, wherein four ELISA positive antibodies, namely R5, R7, R8 and R9, have higher neutralizing activity in all. The rabies virus resistant neutralizing antibody with high affinity, which is provided by the invention, can be used for substituting ERIG and HRIG to carry out active and/or passive immune therapy on rabies virus seriously-exposed persons.

The invention relates to a method for producing rabies vaccines by applying a bioreactor and a sheet carrier, belonging to the technical field of biological products. The method comprises the following steps of: culturing cells by taking BioNOC IITM cell culture disks of the Taiwan SaiYu cell technology Co., Ltd as carriers in a Celligen 310 type 14 L bioreactor with a fixed bed basket-type stirring system; preparing rabies viruses with high titer through specific jar cell density, added viruses and the set parameters of the Celligen 310 type 14 L bioreactor; and producing the rabies vaccines. The method disclosed by the invention has the advantages of high density of cultured cells, high titer of the rabies viruses, high yield and low production cost; the Celligen 310 type 14 L bioreactor is enlarged in surface area used for cell adherence growth by adding the BioNOC IITM cell culture disks and then achieves the cell culture density at 5*108/ml by creating good culture conditions; according to the method disclosed by the invention, the titer of the rabies viruses is correspondingly, greatly logarithmically enhanced and can maximally reach 1*109 FFU/ml, and therefore production cost is greatly reduced; in addition, the method disclosed by the invention is suitable for the large-scale production of rabies vaccines.

The invention provides a method for industrially producing an animal rabies vaccine by utilizing a bioreactor, comprising the following steps of: (1) sterilizing a micro-carrier and the bioreactor, adding a cell growth solution, inoculating Vero cells for culturing, inoculating rabies viruses after the cell on the micro-carrier forms a compact single layer, and continuously culturing to propagate the viruses; (2) inoculating for 18h and then continuously harvesting a virus solution; (3) carrying out ultrafiltration concentration and virus inactivation on the harvested virus solution; and (4) purifying and inactivating the viruses through a column chromatography method to prepare the vaccine. In the invention, the high density culturing of the cells is carried out by utilizing a bioreactor micro-carrier culturing technology to produce the animal rabies vaccine. Compared with the traditional rotating bottle production method, the automation control degree is high, the production can be monitored in real time, the labor power and the cost are reduced, the production land is less, the scale is easy to expand, the produced virus titer is high, the difference among batches is less, the product quality is stable, and the side reaction is less.

The invention provides for an immunogenic rabies vaccine comprising a reduced vaccine dose and methods of pre- and post-exposure immunization with a reduced dose. The concentration of rabies vaccine antigen per dose is preferably less than 2.5 IU/mL.

The invention belongs to the field of biological products, in particular to a process for preparing rabies vaccine by culturing human diploid cells WI-38 and MRC-5 (called as human diploid cells as follows) as toxigenic cells through a Celligen310 bioreactor and taking a rabies virus PM strain as a virus seed, wherein the finished vaccine is prepared by comprising the steps of resuscitating human diploid cells, culturing and proliferating the human diploid cells, inoculating the virus seed of the rabies virus PM strain onto the human diploid cells, domesticating, inoculating and culturing virus seeds, collecting virus filtrate, inactivating, purifying, concentrating, adding a protective solution and the like. Verification indexes of the vaccine meet standards of Chinese Pharmacopoeia of 2010 version. The process is characterized in that the human diploid cells are cultured and prepared as matrix cells by using the Celligen310 bioreactor; therefore, exogenous pollution factors and tumorigenicity of animal passage cells can be avoided. As the inoculated virus strain is the rabies virus PM strain in the preparation process of the vaccine, the immune effect is better than that of the traditional vaccine. The vaccine prepared by using the process has the advantages of being high in purity, good in immune effect and high in safety.