142 results about "Polyproteins" patented technology

The invention provides miniaturized lateral flow chromatographic and lateral flow chromatographic microarray devices (LFM). The miniaturization of lateral flow nucleic acid detection achieved by the present invention offers reduced reagent use, femtomole sensitivity, excellent linear dynamic range, and rapid detection. Moreover, the small feature sizes of capture oligonucleotides renders the potential information capacity of the platform comparable to more traditional spotted fluorescence microarrays as well as improving sensitivity. The LFM devices exemplified herein enable analytes to be detected within 10 seconds from the time of sample introduction to the LFM device. Sample volumes may be as low as about 10 microliters, significantly reducing assay costs and ameliorating reagent storage logistics. Additionally, the miniaturization of lateral flow opens the door to highly multiplexed assays, allowing many proteins or nucleic acids to be detected in a single assay.

Disclosed are useful constructs and methods for the expression of proteins using primary translation products that are processed within a recombinant host cell. Constructs comprising a single open reading frame (sORF) are described for protein expression including expression of multiple polypeptides. A primary translation product (a pro-protein or a polyprotein) contains polypeptides such as inteins or hedgehog family auto-processing domains, or variants thereof, inserted in frame between multiple protein subunits of interest. The primary product can also contain cleavage sequences such as other proteolytic cleavage or protease recognition sites, or signal peptides which contain recognition sequences for signal peptidases, separating at least two of the multiple protein subunits. The sequences of the inserted auto-processing polypeptides or cleavage sites can be manipulated to enhance the efficiency of expression of the separate multiple protein subunits. Also disclosed are independent aspects of conducting efficient expression, secretion, and / or multimeric assembly of proteins such as immunoglobulins. Where the polyprotein contains immunoglobulin heavy and light chain segments or fragments capable of antigen recognition, in an embodiment a selectable stoichiometric ratio is at least two copies of a light chain segment per heavy chain segment, with the result that the production of properly folded and assembled functional antibody is made. Modified signal peptides, including such from immunoglobulin light chains, are described.

Methods and constructs for the introduction of multiple genes into plants using a single transformation event are described. Constructs contain a single 5′ promoter operably linked to DNA encoding a modified intein splicing unit. The splicing unit is expressed as a polyprotein and consists of a first protein fused to an intein fused to a second protein. The splicing unit has been engineered to promote excision of all non-essential components in the polyprotein but prevent the ligation reactions normally associated with protein splicing. Additional genetic elements encoding inteins and additional proteins can be fused in frame to the 5′-terminus of the coding region for the second protein to form a construct for expression of more than two proteins. A single 3′ termination sequence, such as a polyadenylation sequence when the construct is to be expressed in eucaryotic cells, follows the last coding sequence. These methods and constructs are particularly useful for creating plants with stacked input traits, illustrated by glyphosate tolerant plants producing BT toxin, and / or value added products, illustrated by the production of polyhydroxyalkanoates in plants.

An immunogenic or vaccine composition to induce an immune response or protective immune response against West Nile virus (WNV) in an animal susceptible to WNV. The composition includes a pharmaceutically or veterinarily acceptable vehicle or excipient, and a vector. The vector contains heterologous nucleic acid molecule(s), expresses in vivo in the animal WNV antigen, immunogen or epitope thereof, e.g., WNV E; WNV prM and E; WNV M and E; WNV prM, WNV M and E, WNV polyprotein prM-E, WNV polyprotein M-E, or WNV polyprotein prM-M-E. The composition can contain an adjuvant, such as carbomer. Methods for making and using such a composition, including prime-boost regimes and including as to differential diagnosis, are also contemplated.

Compositions and methods for preventing, treating and detecting leishmaniasis are disclosed. The compositions generally comprise fusion polypeptides comprising multiple Leishmania antigens, in particular, KMP11, SMT, A2 and / or CBP, or immunogenic portions or variants thereof, as well as polynucleotides encoding such fusion polypeptides.

HCV variants are described. The variants include polynucleotides comprising non-naturally occurring HCV sequences and HCV variants that have a transfection efficiency and ability to survive subpassage greater than HCV that have wild-type polyprotein coding regions. Expression vectors comprising the above polynucleotides and HCV variants are also described, as are the provision of cells and host cells comprising the expression vectors. Methods for identifying a cell line that is permissive for infection with HCV are also provided, as are vaccines comprising the above polynucleotides in a pharmaceutically acceptable carrier. Additionally, methods for inducing immunoprotection to HCV in a primate are described, as are methods for testing a compound for inhibiting HCV replication.

The invention relates to an immunochromatographic test strip for semi-quantitatively and simultaneously detecting cTnI and Myo and a preparation method thereof. The test strip comprises a base plate, a nitrocellulose membrane, a first bonding pad, a second bonding pad, a sample pad and a water absorbing pad. According to the preparation method, the pretreated nitrocellulose membrane, first bonding pad, second bonding pad, sample pad and water absorbing pad are sequentially and mutually staggered and attached to the base plate so as to obtain the immunochromatographic test strip for simultaneously detecting the cTnI and the Myo. The immunochromatographic test strip disclosed by the invention can be used for simultaneously detecting two proteins with greater abundance difference, providing great convenience for quick diagnosis of clinical myocardial infarction and overcoming and making up the defects of low sensitiveness and narrow linear range during detection of polyprotein in a traditional immunochromatographic test strip technology. The preparation method has the advantages of simple process, low cost, simpleness and convenience in operation and favorable application prospect.

The invention provides HCV fusion proteins that include a mutated NS3 protease domain, fused to at least one other HCV epitope derived from another region of the HCV polyprotein. The fusions can be used in methods of stimulating a cellular immune response to HCV, such as activating hepatitis C virus (HCV)-specific T cells, including CD4+ and CD8+ T cells. The method can be used in model systems to develop HCV-specific immunogenic compositions, as well as to immunize a mammal against HCV.

Methods and constructs for the introduction of multiple genes into plants using a single transformation event are described. Constructs contain a single 5′ promoter operably linked to DNA encoding a modified intein splicing unit. The splicing unit is expressed as a polyprotein and consists of a first protein fused to an intein fused to a second protein. The splicing unit has been engineered to promote excision of all non-essential components in the polyprotein but prevent the ligation reactions normally associated with protein splicing. Additional genetic elements encoding inteins and additional proteins can be fused in frame to the 5′-terminus of the coding region for the second protein to form a construct for expression of more than two proteins. A single 3′ termination sequence, follows the last coding sequence. These methods and constructs are particularly useful for creating plants with stacked input traits and / or value added products.

The invention belongs to great improvement of thick wine brewing technique in food field. The wine is refinedly brewed by taking natural and green glutinous millet, red grapes, mold culture, malt and the like as main materials, adding natural auxiliary material and spices and adopting modern technology. The raw materials can be selected from six kinds of grain products, fruits, flowers and plants, fermentation products, mixing agents and additives. The thick plant wine contains high nutrition, high thermal energy, multiple elements, polyprotein, polysaccharide and multiple amino acids and is a low-fat and low-proof health nourishing drink. The wine is safe and reliable when being drunk for a long time. The thick wine has faint scent and romantic charm, is sweet and clean, and is beneficial to refreshing and restoring consciousness. The thick wine has the advantages of convenient use, being clean and being easy to store and the like, and is especially suitable for middle aged people, old people, women and people who live and work in high cold regions. Being drunk for a long time, the wine is beneficial to bone and fitness strengthening, wind expelling and cold dispersing, qi and blood smoothing as well as longevity promoting, has no damage on human health, contains no melamine, Sudan red or harmful substances, and is coloured naturally. In addition, the prior art and devices are not required to be replaced in transformation of wine brewing enterprise.

Compositions and methods for preventing, treating and detecting leishmaniasis are disclosed. The compositions generally comprise fusion polypeptides comprising multiple Leishmania antigens, in particular, KMP11, SMT, A2 and / or CBP, or immunogenic portions or variants thereof, as well as polynucleotides encoding such fusion polypeptides.

The present invention relates to the field of brain injuries. More specifically, the present invention provides methods and compositions useful in the diagnosis / prognosis / assessment of brain injuries. In a specific embodiment, a method for identifying which patients with traumatic brain injury (TBI) require a head computerized tomography (CT) scan for diagnosing acute intracranial pathology comprises the steps of (a) obtaining or collecting a sample from the patient; (b) measuring the levels of one or more biomarkers in the blood sample obtained from the patient, wherein the biomarkers comprise glial fibrillary acidic protein (GFAP), S100B, metallothionein 3 (MT3), neuron specific enolase (NSE) and intracellular adhesion molecule 5 (ICAM5); and (c) identifying the patient as requiring or not requiring a head CT scan based on the measured levels of one or more of biomarkers comprising GFAP, S100B, MT3, NSE and ICAM5.

An immunogenic or vaccine composition to induce an immune response or protective immune response against West Nile virus (WNV) in an animal susceptible to WNV. The composition includes a pharmaceutically or veterinarily acceptable vehicle or excipient, and a vector. The vector contains heterologous nucleic acid molecule(s), expresses in vivo in the animal WNV antigen, immunogen or epitope thereof, e.g., WNV E; WNV prM and E; WNV M and E; WNV prM, WNV M and E, WNV polyprotein prM-E, WNV polyprotein M-E, or WNV polyprotein prM-M-E. The composition can contain an adjuvant, such as carbomer. Methods for making and using such a composition, including prime-boost regimes and including as to differential diagnosis, are also contemplated.

Synthesis of many proteins is tightly controlled at the level of translation and plays an essential role in fundamental processes such as cell growth and proliferation, signaling, differentiation or death. Methods that allow imaging and identification of nascent proteins allow for dissecting regulation of translation, both spatially and temporally, including in whole organisms. Described herein are robust chemical methods for imaging and affinity-purifying nascent polypeptides in cells and in animals, based on puromycin analogs. Puromycin analogs of the present invention form covalent conjugates with nascent polypeptide chains, which are rapidly turned over by the proteasome and can be visualized and specifically captured by a bioorthogonal reaction (e.g., [3+2]cycloaddition). The methods of the present invention have broad applicability for imaging protein synthesis and for identifying proteins synthesized under various physiological and pathological conditions in vivo.

The invention relates to a method for detecting novel coronavirus 2019-nCoV through multi-protein joint inspection combination, which belongs to the technical field of novel coronavirus 2019-nCoV detection. The invention provides the method for detecting the novel coronavirus 2019-nCoV through multi-protein joint inspection combination. A detection test strip for detecting the novel coronavirus 2019-nCoV antibody is prepared by directly scribing a high-purity and specific novel coronavirus antigen on a nitrocellulose membrane, and IgG, IgM and / or IgA antibodies in a sample can be simultaneously detected by utilizing multiple antigens. Through joint inspection of three or four specific antigens, protein sequences of the antigens are optimally designed, so that the sensitivity and specificity are effectively improved, and the problems of false negative and false positive are effectively avoided.

The invention provides truncated HCV NS5 polypeptides and fusion proteins comprising the truncated NS5 polypeptides, fused to at least one other HCV epitope derived from another region of the HCV polyprotein. The fusions can be used in methods of stimulating an immune response to HCV, for example a cellular immune response to HCV, such as activating hepatitis C virus (HCV)-specific T cells, including CD4+ and CD8+ T cells. The method can be used in model systems to develop HCV-specific immunogenic compositions, as well as to immunize a mammal against HCV.

Disclosed are useful constructs and methods for the expression of proteins using primary translation products that are processed within a recombinant host cell. Constructs comprising a single open reading frame (sORF) are described for protein expression including expression of multiple polypeptides. A primary translation product (a pro-protein or a polyprotein) contains polypeptides such as inteins or hedgehog family auto-processing domains, or variants thereof, inserted in frame between multiple protein subunits of interest. The primary product can also contain cleavage sequences such as other proteolytic cleavage or protease recognition sites, or signal peptides which contain recognition sequences for signal peptidases, separating at least two of the multiple protein subunits. The sequences of the inserted auto-processing polypeptides or cleavage sites can be manipulated to enhance the efficiency of expression of the separate multiple protein subunits. Also disclosed are independent aspects of conducting efficient expression, secretion, and / or multimeric assembly of proteins such as immunoglobulins. Where the polyprotein contains immunoglobulin heavy and light chain segments or fragments capable of antigen recognition, in an embodiment a selectable stoichiometric ratio is at least two copies of a light chain segment per heavy chain segment, with the result that the production of properly folded and assembled functional antibody is made. Modified signal peptides, including such from immunoglobulin light chains, are described.

A foot and mouth disease virus (FMDV) vaccine and method for producing same is described wherein the N terminal portion of the FMDV polyprotein, encoding the four structural proteins, 1A, 1B, 1C, and 1D, are each separated by a non-FMDV protease, preferably a cellular protease, for example, furin. The expression system may be transformed into a cell expressing the non-FMDV protease and the resulting particles recovered for use as a vaccine

An HCV antigen / antibody / antigen assay is provided. The assay employs an isolated first antigen from a region of the HCV polyprotein, and an HCV multiple epitope fusion antigen that includes an epitope from the same region of the polyprotein as the first antigen. Both the first antigen and the multiple epitope fusion antigen bind antibodies present in an HCV-infected sample.

The invention provides a new filaggrinogen or fibrous polyprotein generation accelerator, mantellum protein generation accelerator, cholesterol generation accelerator and cell-activating agent from natural product, which use sap of Betula platyphylla Sukatchev var.japonica Hara as effective constituents.

The present invention relates generally to an immunogenic complex comprising a charged organic carrier and a charged antigen and, more particularly, a negatively charged organic carrier and a positively charged antigen, wherein the charged antigen is a polyprotein of Hepatitis C Virus (HCV), particularly the core protein of HCV, or a fragment thereof, or a fusion protein comprising the polyprotein or a fragment thereof. The complexes of the present invention are useful, inter alia, in vaccine compositions as therapeutic and / or prophylactic agents for facilitating the induction of immune responses, and in particular a cytotoxic T-lymphocyte response, in the treatment of a disease condition which results from an HCV infection.

Methods and constructs for the introduction of multiple genes into plants using a single transformation event are described. Constructs contain a single 5′ promoter operably linked to DNA encoding a modified intein splicing unit. The splicing unit is expressed as a polyprotein and consists of a first protein fused to an intein fused to a second protein. The splicing unit has been engineered to promote excision of all non-essential components in the polyprotein but prevent the ligation reactions normally associated with protein splicing. Additional genetic elements encoding inteins and additional proteins can be fused in frame to the 5′-terminus of the coding region for the second protein to form a construct for expression of more than two proteins. A single 3′ termination sequence, such as a polyadenylation sequence when the construct is to be expressed in eucaryotic cells, follows the last coding sequence. These methods and constructs are particularly useful for creating plants with stacked input traits, illustrated by glyphosate tolerant plants producing BT toxin, and / or value added products, illustrated by the production of polyhydroxyalkanoates in plants.

An immunogenic or vaccine composition to induce an immune response or protective immune response against West Nile virus (WNV) in an animal susceptible to WNV. The composition includes a pharmaceutically or veterinarily acceptable vehicle or excipient, and a vector. The vector contains heterologous nucleic acid molecule(s), expresses in vivo in the animal WNV antigen, immunogen or epitope thereof, e.g., WNV E; WNV prM and E; WNV M and E; WNV prM, WNV M and E, WNV polyprotein prM-E, WNV polyprotein M-E, or WNV polyprotein prM-M-E. The composition can contain an adjuvant, such as carbomer. Methods for making and using such a composition, including prime-boost regimes and including as to differential diagnosis, are also contemplated.

The invention relates to a mathematical model and method for predicting the probability of post-operation recent relapse and metastasis of the giant liver caner of a patient. The mathematical model can be represented through the equation: P=1 / (1+Y), wherein P is the probability of the post-operation recent relapse and metastasis of the giant liver caner of the patient; when P represents the probability of relapse and metastasis occurring in six mouths, Y=exp(4.092+2.073*X12-2.719*X14-2.58*X17+3.039*X19); when P represents the probability of relapse and metastasis occurring in six mouths to twelve months, Y=exp(2.528+1.633*X8+0.971*X12-1.517*X17). The mathematical model and method for predicting the probability of post-operation recent relapse and metastasis of the giant liver caner of the patient mainly have the advantages that multiple indexes relevant to recent relapse and metastasis of the giant liver cancer are comprehensively analyzed based on multiple factors and polyprotein, the effect of predicting post-operation recent relapse and metastasis is achieved through the predicting model, and the model and method have great significance in clinical practice and individualized selection of therapeutic schedules.

The present invention provides a novel cell-based drug screening model of targeting coronavirus protease. In the processing products of the coronavirus polyprotein 1a(1ab), nsp3, nsp4 and nsp6 contain transmembrane (TM) domains which can mediate the protein to locate on the cell endoplasmic reticulum (ER) membrane. Using the characteristics that nsp3, nsp4 and nsp6 contain transmembrane domains and the protein is located in the cell, the present invention constructs a eukaryotic expression DNA construct by secretion reporter gene, wherein the eukaryotic expression DNA construct consists of coronavirus proteases - protease recognition sites - secretion reporter genes. After cell transfection, the expression products of the active releasing reporter gene are cut by the coronavirus protease and are secreted into the cellular supernatant; the activity of the coronavirus protease is detected by the activity of the reporter genes in the cellular supernatant and is used for coronavirus drug screening.

The invention provides, inter alia, methods, devices and reagents for the preparation of native and non-denatured biomolecules using solid-phase extraction channels. The invention is particularly suited for the purification, concentration and / or analysis of protein analytes. The invention further provides, inter alia, methods, devices and reagents for the purification, concentration and / or analysis of multi-protein complexes.

An oral pharmaceutical composition for targeted transport of a platinum complex into the colorectal region, includes a mixture of platinum complex of general formula Iwherein A each independently is an —NH3 group or an amino group containing 1 to 18 carbon atoms, B each independently is a halogen atom, a hydroxy group or a —O—C(O)—R group wherein R each independently is hydrogen atom or an alkyl, alkenyl, aryl, aralkyl, alkylamino or alkoxy group containing 1 to 10 carbon atoms or functional derivatives of these groups, and X each independently is a halogen atom or a monocarboxylate group containing 1 to 20 carbon atoms, or X together form a dicarboxylate group containing 2 to 20 carbon atoms, and at least one excipient selected from the group including a saccharide, oligosaccharide, polysaccharide, modified polysaccharide mucopolysaccharide, protein, oligoprotein, polyprotein, mucoprotein, peptide, oligopeptide and polypeptide, and optionally a lubricant and / or disintegrant, which mixture is optionally compressed into a tablet or contained in a capsule, and this tablet or capsule is optionally coated with a biodegradable layer and / or an outer pH-sensitive colonic layer, and a method of manufacturing and using the composition for treatment of colorectal carcinoma.

The invention provides a double fluorescent quantitative RT-PCR (reverse transcription-polymerase chain reaction) kit for detecting dengue virus and zika virus. The kit comprises a quantitative RT-PCR reaction liquid, an enzyme mixed liquid, a primer probe mixed liquid, standard products (DENV and ZIKV), strong-positive reference products (DENV and ZIKV), weak-positive reference products (DENV and ZIKV), and negative reference products. The kit has the advantages that primer probes with high specificity for DENV and ZIKV are designed according to high conservancy of DENV and ZIKV and the polyprotein protein area with smaller difference between types; the dengue virus and the zika virus can be simultaneously detected through one-time reaction in a single tube by a single tube one-step type double real-time fluorescent quantitative RT-PCR; the specificity and sensitivity are high, and the kit can be used for the laboratory emergency diagnosis of epidemic outbreak caused by dengue virus and zika virus, quick screening, clinical diagnosis, and study of epidemiology of fever eruption virus.

The invention discloses a DNA (deoxyribonucleic acid) vaccine for expressing infectious bursal disease virus polyprotein gene VP243, as well as a construction method and application thereof. A polyprotein gene (VP243) of an infectious bursal disease virus virulent strain (HLJ0504) is optimized by chicken bias codons, and cloned to a eukaryotic expression vector pCAGGS so as to construct a recombinant expression vector pCAGoptiVP243. An initially immunized 14-day SPF chick is injected with 100mu g of the prepared pCAGoptiVP243 though two points of crureus in, a 28-day chick is subjected to the secondary immunization in same dosage and way, and IBDV (infectious bursal disease virus) virulent strain (HLJ0504) strain is attacked in 14 days after secondary immunization. Results indicate that the DNA vaccine can be used for inducing antibody with higher level and providing immunized protection to the virulent strain.