48 results about "Ifn alpha" patented technology

The invention discloses a method for preparing porcine recombinant IFN-alpha. The method is to use a special primer - PCR to amplify target genes of the porcine IFN-alpha, to connect the target genes with an expression vector - pET-28a and an expression vector - pGEX-5X, to transform colon bacillus, to make the colon bacillus highly efficiently express the porcine IFN-alpha and to prepare porcine recombinant IFN-alpha freeze-dried injection through the techniques such as high purification, sterilization, split charging, freeze-drying and so on. The method is shown to have obvious treatment effect on porcine virus diarrhea, influenza and so on and be superior to similar products in the aspects of safety and cure rate through safety experiment, IFN efficacy inspection and clinic probation. The preparation technique is simple; industrialized production is easy; transportation and preservation are easy after freeze-drying. After the porcine recombinant IFN-alpha is injected into a porcine body, the porcine recombinant IFN-alpha is characterized by good healing effect, reliable assembly, no side effect like inocuity and so on, can be used for curing diseases like the porcine virus diarrhea, the influenza and so on, and is novel high-efficient safe nonpersistent broad-spectrum antiviral preparation.

The invention discloses a recombinant porcine alpha interferon, prepared by the following methods: A1, synthesizing an artificially modified porcine alpha interferon; A2, constructing a recombinant eukaryotic expression vector pGAPZ alpha-IFN alpha; and A3, highly expressing IFN alpha by an eukaryotic cell yeast expression system. The invention is characterized by taking a supernatant to purifying the porcine alpha interferon which is cultured by a lot of engineering bacteria and expressed with a chromatography column, then collecting a target protein eluate, filtering through a 0.22 mum microfiltration membrane, then determining the activity by cytopathic inhibition, and calculating the potency unit according to 50% pathology; wherein the cell used for determination is Madin-Darby bovine kidney (MDBK), and vesicular stomatitis virus (VSV) is used for attacking the virus. According to the invention, the recombinant porcine alpha interferon obtained by purification is applied in controlling PCMV, and experiments prove that the recombinant porcine alpha interferon has obvious protection effect on PCMV infected cells. The route of administration of the porcine alpha interferon is injection or mucous membrane administration, and the dosage form comprises injection or nasal drops.

The invention relates to the technical field of biology, in particular to a mesenchymal stem cell culture solution. The mesenchymal stem cell culture solution is prepared from the following components: human platelet lysate, mycillin (Pen Strep), long-acting glutamine, a chemotactic factor XCL1, a chemotactic factor CCL3, a heat shock protein (HSP70), a telomerase inhibitor IFN-alpha 2b and a basal culture medium DMEM. The components of the stem cell culture solution are wide in source, cultured mesenchymal stem cells are high in purity, quick in proliferation and good in dryness, and the mesenchymal stem cell culture solution is suitable for extracorporeal large-scale culture to conduct preclinical study and related clinical study.

The invention relates to an IFN-gama receptor gene (B8R) deleted attenuating carrier of Tiantan recombinant t vaccinia virus as well as a Tiantan recombinant t vaccinia virus AIDS vaccinum deleting IFN-gama receptor gene (B8R) and expressing various HIV-1 antigens, which is constructed based on this carrier, a Tiantan recombinant t vaccinia virus VTKgpe, recombinant t vaccinia virus VTKgpe CGMCC.No.1099 and another a hepatitis-B virus HBSAg antigen, and a hepatitis-B vaccinum of Tiantan recombinant t vaccinia virus of IL-2.

The invention provides a humanized anti-human interferon (IFN) alpha antibody, as well as a coding gene and an application thereof. By combining and applying gene engineering means and phage surface display technology, a plurality of subtype anti-human IFN alpha gene engineering single chain antibodies are screened from a fully synthetic single chain human antibody library, and a gene sequence of a variable region of the antibodies is obtained, holistic monoclonal Fab and IgG antibody recombinant carrier are constructed on that basis, and a high-purity antibody molecule is obtained by expression and purification of mammalian cells. The affinity of combination of the obtained antibody Fab and human IFN alpha-2 is not more than 0.5nM, and the affinity of combination of the obtained antibody Fab and IFN alpha-2 is not more than 5nM. The antibody can be used for effectively blocking the biological activity of IFN alpha-2 and other multiple subtype IFN alpha at a cellular level. The humanized anti-human IFN alpha antibody provides a specific antibody medicine for treatment of IFN related diseases, such as systemic lupus erythematosus and other autoimmunity diseases.

The invention discloses a pig alpha-interferon recombinant adenovirus and constructing method and application to prepare antiviral vaccine, which comprises the following steps: augmentating pig alpha-interferon IFN-alpha gene; cloning; constructing recombinant shuttle vector with IFN-alpha gene; establishing recombinant adenovirus plasmid carrier; obtaining recombinant adenovirus.

The invention refers to the constructive key technology, construction result and the purifying method for producing a-2b interferon of yeast project bacterial strain IFN a-2b/pGAPZa-A/GS115 which can excrete a-2b interferon. The invention uses yeast secretion expressing system as a substitute for colibacillus expressing system, the purpose albumen a-2b interferon needn't repetitiveness, the biological rate activity reaches to 1.0X10 to the 9th power unit/mg albumen, it has no side or noxious effect, and the purification can reach to 95%.

The invention discloses a recombinant interferon--BCG Vaccine bacterial strain and its preparing method, transferring the composed pshuttle phIFN-alpha-2B into the BCG to compose Rbcg-hIFN-alpha-2B bacterial strain. Because it can secrete the interferon IFN-alpha-2B, it can reduce the usage of BCG as compared with that of the wild BCG on the condition of reaching the same and even increased immune effect, thus reducing toxic and side effect; and it can secrete cell factors at the properest time and part. Therefore, it lays the foundation of application to curing cystic tumors.

The invention relates to an application of forsythiaside in anti-PCV2 and anti-PRRSV. The measured prevention effect of the forsythiaside in anti-PCV2 and anti-PRRSV is greater than the treatment effect and further greater than the direct killing effect. The IFN-Alpha secretion after being acted in cells is gradually increased along with the increase of the concentration of the forsythiaside within the range of the safe concentration of the forsythiaside, the virus proliferation is gradually decreased, while the virus proliferation and the amount of the IFN-Alpha show negative correlation. The forsythiaside can induce the IFN-Alpha to express and play the anti-virus role. The invention provides a new method for researching anti-virus traditional Chinese medicine, provides a drug for treating PCV2 and PRSSV and lays the foundation for the action mechanism of anti-PCV2 and anti-PRRSV in vitro of the forsythiaside. The application has the advantages that: materials are easy to obtain, cells are a cell system which can be passaged, the application has no individual difference of animals, and the obtained experimental data is comparatively reliable.

An antiviral drug combination for livestock in the invention relates to an antiviral agent, in particular to an antiviral agent for poultry and livestock. The drug combination is composed of cyanovirin and animal IFN-alpha in mass ratio of 3:1. The invention initially applies cyanovirin to treating viral infection of poultry and livestock, and animal IFN-alpha which is prepared by gene recombination, is contained in species such as pig, chicken, cattle and the like and has high valence, high purity and antiviral effect and cyanovirin with direct antiviral effect are initially applied in a combining way; meanwhile the effect of cyanovirin is direct and rapid and the antiviral effect of IFN-alpha is wide and permanent, and the effect is better than that of single use of cyanovirin or IFN-alpha.

The invention provides a recombination porcine IFN (interferon) alpha1-Fc fusion protein as well as a coding gene and expression, purification and inclusion body renaturation methods of the recombination porcine IFN alpha1-Fc fusion protein. IFN alpha1 is the most common interferon and has remarkable anti-virus, anti-tumor, hematopoietic cell proliferation inhibition, immune adjustment functions and the like. But natural IFN alpha1 is expressed in an organism in a very small amount, lots of IFN alpha1 is difficult to directly extract in vivo for clinical research and application, and the defect of fast clearing speed of the IFN alpha 1 in blood plasma exists. Therefore, the invention provides the recombination porcine IFN alpha1-Fc fusion protein suitable for an escherichia coli prokaryotic expression system, wherein a porcine IFN alpha1 part is an entire sequence of a porcine IFN alpha1 extracellular region, an Fc fragment part comprises a hinge region, a CH2 region and a CH3 region of an antibody, and the porcine IFN alpha1 part and the Fc fragment part are directly fused. The fusion protein provided by the invention maintains the most of biological activity of the IFN alpha 1, the half-life period of the fusion protein is greatly prolonged, and conditions are provided for the industrialization of the fusion protein.

Belonging to the technical field of genetic engineering antibodies, the invention specifically discloses a human vascular endothelial growth factor receptor (VEGFR2 or KDR) targeted antibody JZB00 and a fusion antibody JZA02 of an interferon alpha (IFN alpha) mutant, a preparation method and application thereof. The invention also discloses amino acid sequences of the heavy chain and light chain immunoglobulin molecules of JZA02. The invention also provides a construction method of JZA02 heavy chain and light chain gene, CHO cells are transfected, monoclone is selected by means of limiting dilution method, and then eukaryotic cell secretory expression and affinity chromatography purification are carried out to obtain the fusion antobody. The fusion antibody provided by the invention can specifically bound to VEGFR2, and can inhibit tumor angiogenesis, also a coupling mutation interferon part can play a direct part of tumor killing and immunoregulation, and no toxic or side effect is generated, thus better inhibiting tumor growth. The fusion antibody can inhibit human umbilical vein endothelial cells (HUVEC) in vitro and the proliferation of part of tumor cells, and the inhibition degree is higher than that of a fusion antibody JZA01 of wild type IFN alpha and JZB00.

The invention relates to a chimeric antigen receptor and a preparation method thereof, an NK cell modified by the chimeric antigen receptor and application in treating HBV infection. The chimeric antigen receptor comprises a leader nucleotide artificial sequence, an HBsAg single-chain antibody nucleotide artificial sequence, a CD8alpha hinge region nucleotide artificial sequence, an NKG2D transmembrane region nucleotide artificial sequence, a 2B4 co-stimulation region nucleotide artificial sequence, a CD3zeta signal transduction region nucleotide artificial sequence, a T2A self-cleavable region nucleotide artificial sequence, an IFN-alpha nucleotide artificial sequence and a RQR8 molecular switch region nucleotide artificial sequence. According to the chimeric antigen receptor and the preparation method thereof, the NK cell modified by the chimeric antigen receptor and the application in treating HBV infection, for a common alpha determinant specific antibody, HBsAg in different subtypes can be identified, therefore, the Anti-HBsAg-IFN-alpha-CARNK cell has therapeutic effects on all HBV subtype infection, and meanwhile, the therapeutic dosage and toxic and side effects are also reduced.

The invention discloses an interferon-thymosin broad-spectrum antiviral pharmaceutical preparation for pigs and a preparation method thereof. According to the molecular structure characteristics of interferon alpha 1 (IFN-alpha 1) and thymosin alpha 1 (THY-alpha 1) and the preference of Escherichia coli codon, primers are designed; the PCR method is used for acquiring the fusion gene of the IFN-alpha 1 and the THY-alpha 1; the gene is cloned to a pGEX4T-2 prokaryotic expression vector to construct pGEX4T-IFN alpha 1-THY alpha 1 prokaryotic expression plasmids; and the recombinant plasmids are converted into the BL21 (DE3) recipient bacterium to acquire IFN alpha 1-THY alpha 1 fusion protein through IPTG induction, expression and purification. The biological activity of the protein is detected through cytopathy inhibiting experiment and E-rosette forming experiment, and the result shows that the IFN alpha 1-THY alpha 1 fusion protein has double biological activity.

The present invention relates to the key technology and result of constructing engineering yeast strain IFN alpha-1a/pGAPZ alpha-A/GS115 secreting alpha-1a interferon and the purification process of produced interferon. The present invention adopts yeast secretion expressing system, rather than traditional colibacillus inclusion body expression system, and the target protein alpha-1a interferon has no needs of renaturation, biological specific activity of 1.0E9 unit/mg protein and low toxic side effect. The purification process can obtain target protein alpha-1a interferon of 95 % purity without needing expensive monoclonal antibody column for lowering production cost.

The invention discloses polyethylene glycol-modified human interferon IFN-alpha 2 alpha and human antibacterial peptide LL-37 fusion protein. The obtained IFN-alpha 2 alpha and LL-37 fusion protein have IFN-alpha 2 alpha antiviral activity and LL-37 antibacterial activity, at the same time, half-life period of rats is prolonged from 9.3 hours for IFN-alpha 2 alpha by single usage to more than 78 hours.

An invention relates to use of IFN-alpha resisting marker gene in diagnosis of chronic marrow leukemia. High expression of GSTP1 gene is discovered in cells concerned with tolerant strain KT-1/A3R of IFN-alpha by RT-PCR analysis while low expression of GSTP1 gene discovered in cells with sensitive strain KT-1/A3. Also, high expression of GSTP1 discovered about ITN-alpha resisting tolerant strain KT-1/A3R by Western blotting while low expression of GSTP1 discovered about ITN-alpha sensitive strain KY-1/A3, so that GSTP1 gene can be used as marker gene in diagnosis of drug resistance.

A method for providing activated natural killer (NK) cells comprising the steps of administering to a population of cells which includes lymphocytes and monocytes, an effective amount of an NK cell activating cytokine or a NK cell activating flavonoid, wherein said NK cell activating cytokine is not IL-2 or IFN-alpha; and administering a compound effective to inhibit the production or release of hydrogen peroxide selected from the group consisting of histamine, other H2 receptor agonists, and serotonin.

The invention discloses a dual expression vector of immunostimulatory RNA and liver cancer target gene Pim-3 silent RNA, which is a recombinant vector comprising a DNA sequence transcribing shRNA, a DNA sequence transcribing ssRNA, and a pTZU6+1 vector, and has a sequence as shown in seq.12. The dual expression vector of immunostimulatory RNA and liver cancer target gene Pim-3 silent RNA of the invention is used to transfect mouse liver cancer Hepa1-6 cells; after 24 hours, the cells are detected by Sem-qPCR and Western-blot, and it is found that the expression of Pim-3 in Hepa1-6 cells is down-regulated; after the transfection of Hepa1-6 cells for 24 hours, detection finds that the apoptosis of Hepa1-6 cells is obviously increased; the transfection of Hepa1-6 cells can significantly up-regulate the secretion of I-type interferon (IFN-alpha and IFN-beta) in supernatant of Hepa1-6 cells; therefore, the dual expression vector of immunostimulatory RNA and liver cancer target gene Pim-3 silent RNA of the invention is likely to become a new biological means for liver cancer treatment, and has good application prospects.

The invention relates to the technical field of medicines, and in particular relates to use of a compound MEAN represented in the formula (1) in the specification in preparation of a medicine for inhibiting HCV replication, wherein the medicine is a medicine for inhibiting genotype 2 HCV replication; the compound MEAN is combined with IFN-alpha/RBV/ITX 5061 so as to synergistically inhibit genotype 2 HCV replication; and MEAN is capable of inhibiting genotype 2 HCV replication.

The invention relates to a preparation method of a core fragment rIFN (interferon)-alpha1 of recombinant chicken IFN-alpha, which comprises the following steps that: chicken spleen RNA (ribonucleic acid) is extracted, and is subjected to reverse transcription into cDNA (complementary deoxyribonucleic acid), and the cDNA is used as a template; a nested PCR (polymerase chain reaction) method is adopted, firstly, an outer primer capable of amplifying the chicken full-length gene of IFN-alpha is used for the first round of amplification, and then a self-designed inner primer is used for amplifying corresponding subcloned IFN-alpha1 encoding gene to obtain the core fragment rIFN-alpha1 of the recombinant chicken IFN-alpha. For the core fragment rIFN-alpha1 of recombinant chicken IFN-alpha, the detection of the anti-Newcastle disease virus activity of chicken IFN-alpha1 on BHK-21 cells is carried out, indicating that the highest activity unit of the chicken rIFN-alpha1 is up to 4.32*103U.mg <-1>; and the detection of the anti-Newcastle disease virus activity of chicken IFN-alpha1 is carried out by utilizing SPF (specific pathogen free) chicken embryos cultured virus, indicating that the antiviral activity is higher.

The invention provides a fluorescent quantitation PCR kit for detecting chicken immune-related cytokines and application. The fluorescent quantitation PCR kit is used for SYBR Green I fluorescent quantitation PCR of gene transcription levels of chicken's IFN-alpha, IFN-beta, IFN-gamma, IL-1(beta), IL-2 and IL-6 genes. The kit and a detection method provided by the invention are high in sensitivity, high in specificity, wide in linear concentration range, good in repeatability, and simple and convenient to operate, saves time, and can be used for absolute or relative quantitative detection of chicken immune-related cytokines in various sample types.

The invention belongs to the biotechnology field of genetic engineering vaccine, providing a method for screening recombinant vaccinia virus containing exogenous fusion gene HIV/env/IFN alpha-2b. According to the action principle that the epitope of HIV outer membrane protein has neutralizing antibody determinant which affects virus to attack CD4+ tropism after combined with a target cell; recombinant expression plasmid pJ16env/IFN alpha-2b is constructed by using the method; adopting the liposome dye-transfer process and taking wild vaccinia virus as a transformation gene cause the pJ16env/IFN alpha-2b and the wild vaccinia virus to generate homologous recombination; and the HA characteristic is utilized to screen and purify the pJ16env/IFN alpha-2b recombinant vaccinia virus strain so as to lead the env/IFN alpha-2b fusion gene and the vaccinia virus to be co-expressed. A mice experience indicates that co-expression product of the env/IFN alpha-2b fusion gene and the vaccinia virus can improve the humoral immunity and the cellular immunity of a laboratory mouse; and the expressed protein product can be served as the antigen for immunoprophylaxis. The invention provides an important path for obtaining the outer membrane protein of HIV for researching the antigenicity of HIV structural protein.