75results about How to "Stable potency" patented technology

The invention relates to a pig breeding and breathing complex virus (PPRSV) and pig 2-type loop virus (PCV 2) series gene recombination adenovirus and vaccine in the field of high and new biotechnology. It uses PCR technology to clone the Cap protein gene into the plasmid carrier pShuttle-CMV-GP5 by open type reading rule and transforms tobacillus coli BJ5183strain with cage carrier pAdEasy-1 to capture the recombination plasmid; it uses recombination plasmid HEK293-A cell to capture recombination adenovirus and purifies it to express the recombination adenovirus rAd-Cap-GP5 of PRRSV GP5protein and PCV-2 Cap protein.

The present invention relates to pig reproduction and respiratory syndrome virus recombined adenovirus and vaccine, and belongs to the field of biological high-tech. Through RT-PCR process to proliferate whole PRRSV GP5 sequence, cloning the gene sequence to the shuttle vector pShuttle-CMV of adenovirus carrier system, cotransforming colibacillus BJ5183 strain together with the skeleton vector of adenovirus carrier system to obtain recombinant plasmid, transfecting HEK293-A cell to obtain recombinant adenovirus and plaque purification, and RT-PCR and indirect immunofluorescence technique inspection, the recombinant adenovirus rAd-GP5 expressing PRRSV GP5 protein is constituted. The recombinant adenovirus can set ahead the expression of PRRSV GP5 protein and raise the expression amount to simulate the immune protecting reaction of body effectively.

The invention discloses a porcine reproductive and respiratory syndrome divalent recombination adenovirus vaccine and the preparation method thereof. The invention belongs to the technical field of biological vaccine preparation. The vaccine can be prepared by the following steps: a GP5-2A-M fusion protein gene can be constructed by inserting an FMDV2A gene with self craking between PRRSV GP5 and M protein; homologous recombination is carried out on the GP5-2A-M fusion protein gene and adenovirus backbone plasmid pAdEasy-1; recombination adenovirus rAd-GP5-2A-M is prepared by restriction enzyme and HEK-293A cells transfection, and the divalent recombination adenovirus vaccine is prepared by the technology and the steps such as purification, amplification, and the like. After expression, the aggregate protein GP5-2A-M constructed by the invention is self cracked into GP5 and M protein, as well as exerts the viral neutralization of GP5 and the immune function of the M protein; the vaccine has stable titer with the virulent valence being 10<10.43>TCID<50>/1.0ml, as well as has both the duplication characteristic of a routine attenuated vaccine and the safety of an inactivated vaccine; the divalent recombination adenovirus vaccine can be popularized in and applied to the control work of porcine reproductive and respiratory syndrome.

A preparation method of a purified vitelline freezing antibody for chickens infectious bursal disease is characterized in that: high purity antigens are adopted to vaccinate IBDV B87 to SPF chickens, then velvet urine, idiosome, velvet urine membrence are acquired, grinded, filtered and inactivated to antigens, and the steps are: producing immune eggs, collecting high immune egg, separating vitelline, diluting with distilled water, inactivating, extraction, decentering, inactivating, freezing and checking; the eggs is radiated by Co60 to kill viruses and bacterium, and finally froze by novel immuno-enhancer - Astragali Polysaccharoses and preserved under a temperature of 2-8 DEG C. Composite drugs for prevention and therapy are prepared by adopting the vitelline antibody for chickens infectious bursal disease as major components and cooperating with a novel immuno-enhancer, to decrease the jeopardy of the chicken breeding industry, and improve the living level of people and benefit mankind.

The invention relates to the technical field of medicine, in particular to a cryoprotectant and a lyophilization method for a latex coupled antibody. The cryoprotectant comprises BSA (bovine serum albumin), PEG (polyethylene glycol) and PVP (polyvinylpyrrolidone). The cryoprotectant is designed for the latex coupled antibody and can protect the activity of the latex coupled antibody from being damaged in a lyophilization process in combination with the provided lyophilization method, non-specific aggregation in a redissolution process after lyophilization cannot happen, and the antibody titer keeps stable after redissolution. Repeated experiments prove that the deviation is not obvious, the relative deviation is plus or minus 10% or smaller, and the cryoprotectant has good lyophilization protection effect.

The invention discloses a recombinant anine interferon alpha standard substance, a preparation method and potency determination method thereof. BL21/pET-28 alpha-rCaIFN alpha recombinant bacteria of the expression recombinant anine interferon alpha is fermented and induced, and supernatant after bacterial cell disruption is collected directly to extract the total protein; thereafter, a pure product of the preparation is obtained by three-step purification, a freeze-drying protective additive is added in the pure product for vacuum freeze drying, then a standard substance is obtained, wherein the valence is 1.0*1.04 IU/mL, the purity is 97.34% by a SDS-PAGE detection, the purity is 99.79% by a HPLC detection, and the relative molecular weight is 33 KD. According to the preparation methd, the prepared pecombinant anine interferon alpha as a standard substance can be used for comparison of characteristic identification and potency determination of the recombinant anine interferon alpha, when the standard substance is taken as a standard, the determining results of the potency determination of different batches of the recombinant anine interferon alpha are more reliable and more dependable.

The invention relates to an improved body fluid color-developing fungi 1,3-beta-D-glucan detection kit, which comprises a reaction main-agent, a main agent combination solution, a sample treatment fluid, sterile water, a standard substance and a quality control material. With Tachypleus tridentatus Leach or Limulus polyphemus Linnaeus hemocyte lysate as a main raw material, the reaction main-agent contains G factor, B factor, C factor, clottable protein and a polypeptide chromogenic substrate. By a new preparation process, the reaction main-agent has characteristics of high stability and small intra-assay and inter-assay difference. The main agent combination solution adopts a new formula. After the main agent combination solution is mixed with the reaction main-agent, Tachypleus Amebocyte Lysate endotoxin reaction branch can be effectively shielded. According to the kit, rate-method enzyme kinetics detection is carried out by ELIASA. The detection speed is fast, and anti-interference performance is strong. In addition, the preparation technology is simple, and the product has stronger stability and higher specificity and sensitivity.

The invention discloses a lytic vibrio parahaemolyticus bacteriophage RDP-VP-19003 and an application thereof to pathogenic vibrio parahaemolyticus infection of aquatic animals. According to the invention, the bacteriophage is separated from wastewater of Qingdao Jimo aquaculture farmers by using a double-layer plate method, is named as vibrio parahaemolyticus bacteriophage RDP-VP-19003, and has apreservation number of CGMCC No. 19385. The vibrio parahaemolyticus bacteriophage RDP-VP-19003 provided by the invention has relatively high temperature tolerance and a relatively wide acid-base tolerance range, and is beneficial to industrial production and preparation. The bacteriophage has a strong cracking effect on the vibrio parahaemolyticus BV-19004, can effectively prevent and control generation and propagation of the vibrio parahaemolyticus, and can be used for efficiently and reliably treating aquatic animal diseases caused by the vibrio parahaemolyticus and improving the water environment.

A composite medicine containing norfibrase features that a human hemoalbumen a protecting agen is added to form a water film surrounding the norfibrase protein molecule to prevent it from being contacted with solvent (water), that is, from being adsorbed to lower its medical value.

The invention discloses a shuttling intracellular antibody TAT-4F for H3N2-type canine influenza virus. The H3N2 virus is A-type influenza virus, the main neutralizing antibody is from hemagglutinin (HA), therefore, the HA becomes an original main research target; then the influenza virus has high variability, the immune cross reaction among different variable branches is weaker, M1 antibody can be combined with M1 protein to inhibit the activity and disturb multiplication, transcription and release of the influenza virus so as to play a role in resisting the virus; therefore, the M1 protein is selected for preparing the corresponding antibody to obtain stable titer, coupling the antibody with TAT protein PTD (Protein Transduction Domain) capable of transducing biomacromolecules into cellsand expressing fusion protein TAT PTD-M1 ScFv, and then a shuttling antibody for resisting the canine influenza virus is prepared to provide a new path for treating the canine influenza virus.

The invention discloses a porcine parvovirus BQ-C strain and application of the porcine parvovirus BQ-C strain to the preparation of an inactivated porcine parvovirus vaccine, and belongs to the field of biological vaccines. The porcine parvovirus BQ-C strain has the advantages of high immunogenicity, good cultural character and the like; and porcine parvovirus (BQ-C strain) are inoculated to swine testis (ST) cells, a culture is harvested and inactivated by using binary ethylenimine (BEI), and the culture and a commercial MontanideTM ISA15AVG adjuvant of SEPPIC are mixed and emulsified to form the inactivated porcine parvovirus vaccine. The experiment proves that: diseases caused by porcine parvoviruses can be prevented by immunizing primiparous healthy female sows by using the prepared inactivated porcine parvovirus vaccine; and moreover, the invention has the advantages that the vaccine is safe, an antibody can be quickly generated, the immune period is long-lasting, and the like.

The invention relates to the technical field of biology, and particularly provides a fusion protein of an infectious bursal disease virus, a preparation method, application, an expression system of the fusion protein and a vaccine comprising the fusion protein. The fusion protein comprises a VP2 section and a VP3 section, wherein the VP2 section is expressed by a nucleotide sequence shown in SEQ ID NO.1, the VP3 is expressed by a nucleotide sequence shown in SEQ ID NO.2. The fusion protein is obtained by tandem expression of the gene sequences of the VP2 and the VP3 of the infectious bursal disease virus. By analyzing the gene sequences of the VP2 and the VP3 and selecting zones high in antigenicity and easy for high expression to link tandemly, the obtained fusion protein has the advantages of high antigenicity and high expression amount. By adopting the fusion protein as an antigen to prepare the vaccine, the prepared vaccine has the advantages of good safety, stable potency and no toxic and side effects. The invention further provides the preparation method and application of the fusion protein, and the vaccine prepared from the fusion protein.

The invention discloses a shuttling intracellular antibody for H3N2-type canine influenza virus. The H3N2 virus is A-type influenza virus, the main neutralizing antibody is from hemagglutinin (HA), therefore, the HA becomes an original main research target; however the influenza virus has high variability, the immune cross reaction among different variable branches is weaker, M1 antibody can be combined with M1 protein to inhibit the activity and disturb multiplication, transcription and release of the influenza virus so as to play a role in resisting the virus; therefore, the M1 protein is selected for preparing the corresponding antibody to obtain stable titer, coupling the antibody with TAT protein PTD (Protein Transduction Domain) capable of transducing biomacromolecules into cells andexpressing fusion protein TAT PTD-M1 ScFv, and then a shuttling antibody for resisting the canine influenza virus is prepared to provide a new path for treating the canine influenza virus.

The invention provides a hemocoagulase freeze-dried powder injection and a preparation method thereof, and belongs to the field of biological products and preparations. The hemocoagulase freeze-dried powder injection takes hemocoagulase as an active ingredient, and a freeze-dried protective agent consists of gelatin, dextran-20, and sorbierite. The hemocoagulase freeze-dried powder injection can be used as a haemostatic and the like.

The invention discloses a lysozyme hydrochloride toothpaste and a preparation method thereof. The lysozyme hydrochloride toothpaste comprises an active ingredient and auxiliary materials, wherein the active ingredient is lysozyme hydrochloride, and the weight percentage of lysozyme hydrochloride in toothpaste is 0.3-4.6 percent; the auxiliary materials are any acceptable auxiliary materials for preparing the toothpaste, and comprise a humectant, a thickener, a foaming agent, a protective agent, an abrasive, a flavoring agent and other additives. The lysozyme hydrochloride toothpaste has the advantages that the cleaning capability is strong, the mouth feel of tooth brushing is excellent, the effect of treating periodontal diseases, such as inhibiting of dental plaque and/or relieving of gingivitis and periodontitis, are realized, and more safety in use is realized.

The invention discloses a salmonella abortus equi horse derived strain and an application thereof in preparation of a salmonella equina inactivated vaccine, and belongs to the technical field of biological vaccines. The salmonella abortus equi horse derived strain is named as 20180316.H.AES.G and is preserved in the common microorganism center of the horse China General Microbiological Culture Collection Center, wherein the culture collection number is CGMCC No.18341. The salmonella abortus equi horse derived strain 20180316.H.AES.G provided by the invention has the advantages of good immunogenicity, good culture characteristics and the like. An inactivated vaccine is prepared by the steps of inoculating the salmonella abortus equi horse derived strain 20180316.H.AES.G to a salmonella equienrichment broth, performing culturing, and then performing inactivating with 0.2% v/v formaldehyde. Experiments prove that when a mouse is immunized by the prepared inactivated vaccine of the salmonella abortus equi, the mouse can be free from death after challenge, and safety is good after a horse body is immunized. Therefore, the vaccine has the advantages of safety, low-dose immunization andthe like. The invention provides the inactivated vaccine of salmonella abortus equi for the first time, and the inactivated vaccine can be used for preventing abortion caused by salmonella abortus equi.

The invention relates to the technical field of veterinary biological products, in particular to an avian adenovirus 4, 8 and 11 type trivalent vaccine as well as a preparation method and application thereof. The fowl adenovirus trivalent vaccine comprises a fowl adenovirus type 4 vaccine strain QYH2019-YN, a fowl adenovirus type 8 vaccine strain QYH2020-HB and a fowl adenovirus type 11 vaccine strain QYH2020-SY. The invention provides a fowl adenovirus trivalent vaccine which is prepared from a fowl adenovirus type 4 vaccine strain, a fowl adenovirus type 8 vaccine strain and a fowl adenovirus type 11 vaccine strain which are obtained by newest separation, and has an effective prevention effect on fowl adenovirus type 4, fowl adenovirus type 8 and fowl adenovirus type 11 as well as chicken inclusion body hepatitis and hydropericardium-hepatitis syndrome caused by the fowl adenovirus type 4, fowl adenovirus type 8 and fowl adenovirus type 11.

The invention relates to the technical field of veterinary biological products, in particular to a group I avian adenovirus type 8 strain and application thereof. The group I avian adenovirus type 8 strain ZMYTAV-8 strain, with an accession number of CGMCC No. 14297 has the characteristics of high toxin production and good immunogenicity. The strain can effectively prevent chicken inclusion body hepatitis caused by serum avian adenovirus type 8. Therefore, the invention also provides application of the avian adenovirus type 8 strain ZMYTAV-8 in the preparation of drugs for the prevention of inclusion body hepatitis. Further, the present invention provides a bivalent inactivated vaccine of avian adenovirus serotype 4, serotype 8 for the prevention of pericardial effusion-hepatitis syndromeand inclusion body hepatitis. The vaccine has good immunogenicity and can prevent avian diseases caused by adenovirus, such as inclusion body hepatitis and pericardial effusion-hepatitis syndrome, which are popular in recent years. The vaccine has 100% protection rate against local isolates, and has high safety, rapid antibody production and stable efficacy.

The present invention discloses bovine parainfluenza virus PBIV3-B strain and application thereof, the bovine parainfluenza virus PBIV3-B strain has the advantages of good immunogenicity and culture characteristics and the like, the bovine parainfluenza virus PBIV3-B strain is inoculated into MDBK cell to obtain a culture matter, after the culture matter is inactivated with (beta-Propiolactone), the culture matter is mixed with commercial MontanideTMISA15AVG adjuvant of SEPPIC of France for emulsion to obtain a bovine parainfluenza virus inactivated vaccine. Results showed that when the bovine parainfluenza virus inactivated vaccine prepared by the method is used for immunization of a weaned healthy negative cow, diseases caused by the bovine parainfluenza virus type 3 can be prevented, and the bovine parainfluenza virus inactivated vaccine has the advantages of safety, fast antibody production and lasting immunity period and the like.

The invention discloses a nucleic acid and recombinant protein co-immune vaccine based on a classical swine fever virus gene as well as a preparation method and application of the co-immune vaccine. The preparation method comprises the steps that firstly, optimized recombinant plasmids and recombinant classical swine fever vaccine antigen protein are obtained by combining gene engineering and cellengineering; then the nucleic acid sequence and the recombinant protein are compounded into different liquid phase layers of a biphasic adjuvant, so that the obtained co-immune vaccine is good in immunogenicity and short in immune blank period, can rapidly induce an organism to generate high-level, high-affinity and durable antibody response and immune memory, effectively protects a target animalfrom being attacked by classical swine fever virulent viruses, and the method can be applied to preparation of products for preventing/treating classical swine fever.

The invention belongs to the technical field of ruminant nutrient substance rumen protection, and particularly relates to a rumen compound preparation, a preparation method thereof and an application of the preparation, wherein the rumen compound preparation is used for ruminant feed, can improve the use performance of the feed and is high in nutrient substance content and good in thermal-sensitive properties. The rumen compound preparation comprises components A, B, C, D and E, wherein the component A is prepared from, by weight, 11 parts of methionine, 16 parts of lysine, 3 parts of niacin and 10 parts of glycine betaine, the component B is a ruminant enzyme preparation, the component C is a ruminant probiotics, the component D is a vitamin for a ruminant, and the component E is an rumen auxiliary material. The component A, the component B, the component C, the component D and the component E are mixed according to the weight ratio of A:B:C:D:E=2:1:1:1:5 to prepare the rumen compound preparation. The rumen compound preparation has the advantages that various nutrient substances can be simultaneously provided, absorption of the nutrient substances in the feed can be promoted, feed use ratio is increased, and the added nutrient substances are high in use ratio.

The invention provides a construction method of a gene engineering strain of Lactobacillus paracasei with high bacteriocin yield, relates to a construction method of the gene engineering strain of the Lactobacillus paracasei and aims to solve the problem of low bacteriocin yield of the Lactobacillus paracasei at present. The method comprises steps as follows: step one, a prcK gene is amplified from Lactobacillus paracasei HD1.7; step two, a plasmid pMD18-T-prcK is constructed; step three, a recombinant overexpression vector pRSFDuet-1-prcK is constructed; step four, a recombinant overexpression vector pRR-tet is constructed; step five, the overexpression vector pRR-tet is used for performing electrotransformation on Lactobacillus paracasei HD1.7 competent cells; step six, overexpression mutant strains of the prcK gene are screened and identified. The bactericidal capacity of the gene engineering strain is increased by 17.86% compared with that of original strains; bacteriocin titer of fermentation broth after passage is 2372.46 plus or minus 46.27 AU/mL, and the titer after the passage is more stable; the method is applied to bacteriocin production.

The invention belongs to the technical field of biology and discloses swine influenza virus H3N2 subtype HA-1 protein recombinant suipoxvirus and a preparation method thereof. An HA1 gene is designed and synthesized according to the gene sequence of HA1 of a A/swine/Guangxi/1/2004(H3N2) strain of the swine influenza virus, and the gene is cloned in a suipoxvirus vector pUSZ11 to obtain a transfer vector pUSZ11/H3. The recombinant virus is a positive clone obtained by infecting and transfecting PK15 cells with the suipoxvirus and the transfer vector pUSZ11/H3 and performing homologous recombination. The recombinant suipoxvirus can express HA1 protein stably, and after infection with the virus, the cytopathy becomes regular, the toxic effect of the breeding virus is stabilized at 107TCID50/mL, the breeding virus can effectively stimulate the immune protect reaction of organisms and can be used in the preparation of medicines for preventing and/or treating infection with swine influenza virus H3N2 subtype.