48 results about "Polyproteins" patented technology

A chimeric nucleotide sequence that corresponds to an encoded fusion protein comprising a polyepitope resulting from selecting and juxtaposing multiple epitopes from a coronavirus protein to induce an immune response in mammals. In one embodiment, said fusion protein comprises: a) a first peptide consisting of epitopes found in the amino acid sequence of replicase polyprotein 1ab (PR1ab); b) a first spacer; c) a modified form of herpes simplex virus type 1 (HSV-1) glycoprotein D (gD). In one embodiment, the replicase polyprotein is defined by SEQ ID NO: 96 flanked by a gD fragment comprising the amino acid sequence defined by SEQ ID NO: 98 in the N-terminal portion and another gD fragment comprising the amino acid sequence defined by SEQ ID NO: 100 in the C-terminal region. Use of the fusion protein has surprising results in inducing cellular and humoral immune responses against coronavirus, SARS-COV-2, and related viruses.

Disclosed herein are vaccine constructs for producing a virus-like particle (VLP) capable of raising an immune response to an immunogen, and uses thereof, wherein the constructs comprise nucleic acid sequences encoding an immunogen and a polyprotein, wherein the polyprotein comprises two or more viral structural proteins, wherein at least two of the two or more viral structural proteins are separated by a signal peptidase sequence such that, when the polyprotein is expressed in a host cell, the signal peptidase sequence undergoes host cell peptidase-dependent cleavage to liberate the two or more viral structural proteins, thereby allowing the liberated structural proteins to self-assemble into a VLP carrying the immunogen.

Methods of labeling, processing and analyzing protein samples. Surface-accessible residues in a protein sample (such as a proteome) can be labeled using a carbodiimide coupling agent an amine comprising a click chemistry handle, such as glycine propargyl amide. The click chemistry handle can then be derivatized using high-yield "click" chemistry to form a stable linkage with an enrichable label. The proteome can then be digested, peptides that have been modified can be enriched, and the enriched peptides can be analyzed, for example, by mass spectrometry. The analyzed peptides can constitute a highly enriched subset of the original peptides while still representing many sites of modification on many proteins (~95%) originally present in the proteome. The methods can be used to identify solvent-accessible regions of proteins, conformational changes, and drug targets across a whole proteome.

The present disclosure relates to methods, devices and systems for exposing microorganisms or compositions comprising functional proteins to an oscillating electric field substantially free of a magnetic component in order to improve stress responses, survival and metabolite or protein production. In embodiments, a controller drives one or more insulated emitters with a true AC signal or pulsed DC signal, in stepped, swept or shuffled patterns of an electric field oscillating at one or more frequencies in the range 210 Hz to 99 kHz. Exposure to the field provides surmounting energy that enhances protein breathing, catalysis, and fundamental functions of multiprotein complexes including transcription factors and ribosomes, thereby improving stress resistance of microorganisms, their viability during fermentation, drying, storage, transport, and / or increasing production of functional or therapeutic proteins, metabolites or microbial lysates. The technology is applicable to manufacture of probiotics, therapeutics, metabolites, food, alcohol, feed and bioenergy, cosmetics, nutraceuticals and biopharmaceuticals..

The invention relates to the technical field of tag peptides, and particularly discloses a tag peptide combined on the surface of corynebacterium, and the amino acid sequence of the tag peptide is shown as SEQ ID NO: 1 or SEQ ID NO: 2. The tag peptide provided by the invention enriches the types of tag peptides bound to the surfaces of current bacteria, and corynebacterium bacteria bound with the tag peptide have wider adaptability to pH and can be compatible with the optimal activity conditions of more foreign proteins; the molecular weight of the tag peptide is only 3.5 kDa, so that the interference to the structure or function of an exogenous protein can be remarkably reduced; in addition, the tag peptide can be fused at the N end or the C end of the target protein, the binding efficiency is not affected, simultaneous display of multiple proteins is supported, and a basis is provided for multi-enzyme cascade reaction and multivalent vaccine development.

The present invention discloses a gene cluster for replacing the γ-PGA synthase sourced from PgsA and a method for synthesizing polyglutamic acid therefrom. The present invention clones the γ-polyglutamic acid synthase gene cluster pgsBCA from Bacillus licheniformis into a high-glutamic acid-producing strain, Corynebacterium glutamicum F343, for exogenous expression. On this basis, pgsA is replaced with pgsA from other strains, namely Bacillus subtilis, Bacillus methylotrophicus, Bacillus amyloliquefaciens, and Bacillus anthracis, to obtain recombinant strains BCA'(BS), BCA'(BM), BCA'(BAM), and BCA'(BAN). The γ-PGA yields produced by BCA'(BS), BCA'(BM), and BCA'(BAM) increased by 54.73%, 34.87%, and 25.10%, respectively, while the γ-PGA yield produced by BCA'(BAN) decreased by 24.09%. Therefore, replacing the source of PgsA in a specific PgsBCA multiprotein complex is an effective way to further improve γ-PGA production.

The present invention relates to a method for the purification of one or more recombinantly expressed protein(s) of interest (POI(s)), comprising the steps of expressing the POI(s) as a fusion protein in which one or more copies of the POI(s) is / are fused to a protomer of a multiprotein complex (MPC), assembling said MPC, said MPC comprising one or more of said fusion protein(s) and any further protomers that are part of said MPC, wherein one or more of the protomers assembled in the MPC contain(s) at least one histidine-rich amino acid sequence, subjecting said MPC in solution to a multi-metal chelate in solution, thereby forming an affinity precipitate, and purifying said affinity precipitate. The present invention further relates to respective affinity precipitates obtained by said method.

The invention discloses a cell strain capable of stably expressing a multi-protein tag as well as a construction method and application of the cell strain, and belongs to the technical field of biology. The cell strain is transfected by a lentiviral vector, a fusion expression vector containing P2A self-cleavage polypeptide and a plurality of protein tag coding sequences is integrated into a host cell, and a stable expression cell strain and a fusion protein expressing a plurality of protein tags including green fluorescent protein are obtained through antibiotic screening. The cell strain can be used for antibody specificity verification, protein interaction research and multi-label system standardization experiment platforms, and has the advantages of stable expression, simple operation, reliable result and the like.

The present invention relates to a vaccine composition for breaking self-tolerance against a self-protein of a host, in particular for breaking self-tolerance against endogenous cytokines, in particular against the endogenous IL-4, IL-5, IL-13, IL-31 and IL-33 proteins in an animal host. The vaccine composition of the invention contains a polyprotein, a DNA encoding for the polyprotein and / or an RNA encoding for the polyprotein and one or more immunostimulatory oligonucleotides. The polyprotein comprises at least two self-protein segments of the host and one or more T-cell epitopes of non-host origin in between and / or adjacent to the at least two self-protein segments.The present invention further concerns the use of the vaccine composition for the prevention and / or treatment of diseases including the prevention and / or treatment of a pruritic condition and / or an allergic condition. In another aspect, the present invention provides a method for detecting the presence of autoantibodies against self-proteins that can be generated with the vaccine composition of the invention.

The present invention discloses a gene cluster for replacing a γ-PGA synthase sourced from PgsA and a method for synthesizing polyglutamic acid therefrom. The γ-polyglutamic acid synthase gene cluster pgsBCA of Bacillus licheniformis is cloned into a Corynebacterium glutamicum F343 strain with high glutamate production for exogenous expression. On this basis, pgsB is replaced with pgsB from other strains, namely Bacillus subtilis, Bacillus methylotrophicus, Bacillus amyloliquefaciens, and Bacillus anthracis, to obtain recombinant strains B′(BS)CA, B′(BM)CA, B′(BAM)CA, and B′(BAN)CA. The yields of γ-PGA produced by B′(BS)CA, B′(BM)CA, and B′(BAM)CA are increased by 48.00%, 107.87%, and 98.43%, respectively, while the yield of γ-PGA produced by B′(BAM)CA is significantly decreased. Therefore, replacing the source of PgsB in a specific PgsBCA multiprotein complex is an effective way to further improve γ-PGA production.

The invention relates to the field of traditional Chinese medicine hair care, and discloses a multi-protein-containing repairing hair mask and a preparation method thereof. The hair mask is prepared from silk protein hydrolysate, wheat protein peptide, collagen tripeptide and polygonum multiflorum / cacumen biotae / ginseng traditional Chinese medicine extract according to a specific proportion through a step-by-step enzymolysis and low-temperature emulsification process. The multi-protein system can permeate into the deep layer of hair epidermis to repair damaged hair cuticles under the synergism of the active ingredients of the traditional Chinese medicines. Compared with a traditional single-protein hair mask, the product has the advantages of remarkable repair depth and lasting effect, can effectively improve the strength and glossiness of hair, and is suitable for caring hair damaged by ironing and dyeing.

The present disclosure provides dimeric deamination polyproteins comprising a first nucleotide deaminase, a second nucleotide deaminase, a first nuclease-active Cas-alpha 10 polypeptide, and a nuclease-active second Cas-alpha 10 polypeptide. Also disclosed are methods of modifying a double-stranded DNA target site and methods of multiplexed editing using a deamination polyprotein and guide polynucleotides, wherein a dimeric deamination polyprotein and the guide polynucleotide form a complex that recognizes and binds to a dsDNA target site.

A number of protein aggregation diseases are associated with accumulation of misfolded proteins, which are known as protein aggregates, including, but not limited to, neurodegenerative and non-degenerative diseases and disorders. The present disclosure provides an assay, compositions and kits for the qualitative and quantitative assessment of aggregated proteins in solution using a microparticle immunocapture assay that combines the advantages inherent to a specific first and second capture moiety that binds specifically to an aggregated protein which can reveal at the same time the amount and the size of aggregates measured in a sample, fluid, tissue, cavity, or pharmacological product.

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.

The present invention discloses a high-throughput cell imaging method and application based on fluorescent barcoded immunolabeling. A fluorescence barcoded immunolabeling signal amplification method based on the HCR reaction is used to construct ADC complexes corresponding to various proteins in cells. The antibody end recognizes the target, and the DNA end triggers the HCR reporting signal. Multiple proteins in cells are simultaneously incubated and labeled for fluorescence imaging. The artificial + computer image intelligent recognition combination is used for multi-color differentiation of cells to distinguish and quantify multi-protein fluorescence imaging, which is used for multi-protein imaging detection and analysis in vitro tumor cells. The present invention realizes simultaneous imaging of multiple proteins in cells based on fluorescence reporting signals under different combinations, solves problems such as cumbersome operation and cell denaturation caused by repeated imaging, and at the same time combines artificial and image intelligent recognition to achieve the differentiation and quantitative analysis of each protein, which has important application value in the fields of multi-target and multi-omics analysis.

The present disclosure concerns a Protein Quantitation Reporter (PQR) linker which is capable of being cleaved during the translation of a messenger RNA to quantify a protein of interest. The PQR linker can encode a peptide of SEQ ID NO: 23 and have the nucleic acid sequence of SEQ ID NO: 25. The PQR linker is located in a nucleic acid molecule encoding a poly-protein between a reporter protein and the protein of interest. While the messenger RNA encoding the poly-protein is being translated, the presence of the PQR linker causes cleavage of the poly-protein and consequently the release at a stoichiometric ratio of the reporter protein and the protein of interest. The signal associated with the cleaved reporter protein can be measured to estimate or quantify the protein of interest.

The present invention relates to a method for the purification of one or more recombinantly expressed protein(s) of interest (POI(s)), comprising the steps of expressing the POI(s) as a fusion protein in which one or more copies of the POI(s) is / are fused to a protomer of a multiprotein complex (MPC), assembling said MPC, said MPC comprising one or more of said fusion protein(s) and any further protomers that are part of said MPC, wherein one or more of the protomers assembled in the MPC contain(s) at least one histidine-rich amino acid sequence, subjecting said MPC in solution to a multi-metal chelate in solution, thereby forming an affinity precipitate, and purifying said affinity precipitate. The present invention further relates to respective affinity precipitates obtained by said method.

The present disclosure relates to methods for preparing and using multiprotein complexes. The complexes can establish multivalent binding mechanisms for multiple targets in solution or on a surface. The complexes have various applications and can be used, for example, for activation and / or expansion of T cells.

Provided is a multi-protein degradation agent having an imide skeleton, which can degrade multiple proteins comprising c-Myc, GSPT1, CK1α and IKZF (1 / 2 / 3) proteins, and therefore can be used for treating diseases or disorders related thereto, such as cancer.

Provided herein, are compositions based on retroviruses (e.g., lentiviruses) comprising one or more nucleic acid molecules encoding retroviral Pol polyprotein components and a nucleic acid molecule comprising one or more transgene sequences flanked by long terminal repeat sequences, for delivery of the one or more transgenes to a target cell ex vivo or in vivo. The compositions are useful for delivering to a target cell (e.g., hematopoietic stem cells (HSCs), liver cells, ocular cells, muscle cells, epithelial cells, T cells, etc.) and / or stably expressing any transgene (e.g., beta-globin, Factor VIII, RP GTPase regulator (RPGR), dystrophin, cystic fibrosis transmembrane conductance regulator (CFTR), a chimeric antigen receptor, etc.) with a biological effect to treat and / or ameliorate the symptoms associated with any disorder related to gene expression (e.g., sickle cell disease, beta-thalassemia, haemophilia B, retinitis pigmentosa, Duchenne muscular dystrophy, cystic fibrosis, cancer, etc.).

The present invention discloses the application of PHT-427 in the preparation of a drug for inhibiting the activity of the monkeypox core protease. By using high-throughput drug screening technology and through multiple screenings, a small molecule compound PHT-427 that can efficiently inhibit the core protease I7L was obtained. Through experiments, it was confirmed that the IC 50 value of PHT-427 for the monkeypox virus core protease I7L is 9 μM. In vitro enzyme activity experiments demonstrated that the compound PHT-427 can specifically bind to the active site of the monkeypox virus core protease, significantly inhibit the catalytic function of this protease, block the cleavage processing of viral polyproteins and the maturation and assembly of virus particles, and thereby exert an antiviral effect.

The present disclosure provides engineered Class 1 Type I CRISPR-Cas (Cascade) systems that comprise multi-protein effector complexes, nucleoprotein complexes comprising Type I CRISPR-Cas subunit proteins and nucleic acid guides, polynucleotides encoding Type I CRISPR-Cas subunit proteins, and guide polynucleotides. Also, disclosed are methods for making and using the engineered Class 1 Type I CRISPR-Cas systems of the present invention.

The invention relates to the technical field of early tumor screening and molecular diagnosis, in particular to a protein detection marker for screening early gastric cancer, and particularly relates to a mass spectrum-based multi-protein marker detection method. The protein composition shows excellent diagnostic performance in early gastric cancer detection, achieves an AUC value of 0.95 or above in a clinical queue, and has clinical application potential.

The present application provides a kind of based on PBA data's microorganism structure proteomics analysis method, by inputting the PBA original data of microorganism structure, extract the encoding nucleotide fragment containing specific information from the PBA original data, obtain intermediate transmission data;Then extract the protein expression data of the microorganism structure from the intermediate transmission data.From protein expression data, the total protein expression data of sample, the protein combination data of multiple protein co-expression, the subgroup data obtained after clustering analysis based on the protein expression data of the microorganism structure are analyzed to obtain machine learning.In the multi-level of protein expression, protein combination, subgroup, potential biomarker screening can be carried out.The analysis method described in the present application analyzes the total protein expression, protein combination form, microorganism structure subgroup and its protein fingerprint characteristics and quantification on the level of single microorganism structure, fills the blank of bioinformatics analysis in the field of microorganism structure proteomics detection.

The present invention relates to the field of biotechnology, and specifically to the use of the mTOR inhibitor Torin1 in the preparation of drugs for cholestatic bile duct injury. mTOR is an evolutionarily conserved serine / threonine kinase that plays a key role in cell growth and metabolism. Torin 1 is an inhibitor of mTOR. mTOR regulates cell growth, movement and metabolism by forming two multiprotein complexes, mTORC1 and mTORC2. mTORC1 is sensitive to nutrients, and mTORC2 is regulated by PI3K and growth factor signaling pathways. The present invention found that Torin 1 has a significant therapeutic effect on cholestatic bile duct injury. After treatment, it can significantly improve liver function indicators and pathological changes such as inflammatory cell infiltration, bile duct hyperplasia, and periductal liver fibrosis in patients with cholestatic bile duct injury; this inhibitor can significantly reduce the level of inflammatory factors in animal models of cholestatic bile duct injury.

The invention belongs to the technical field of biological medicine, and particularly discloses a multi-protein marker combination for intervertebral disc degeneration prediction and severity evaluation and a machine learning algorithm. The marker combination comprises any three or more of COL6 [alpha] 3, REG1beta, ATF5, CAP1, MAGEA4 and LILRB3, and preferably six proteins are combined for use. And quantitatively detecting the expression level of the protein in the blood plasma through enzyme-linked immunosorbent assay (ELISA), constructing a prediction model in combination with a binary Logistic regression algorithm, and outputting an intervertebral disc degeneration grading result. Experiments show that the AUC (area under curve) of joint detection of six proteins reaches 0.870, which is averagely improved by 11.8% compared with that of single protein detection, and the AUC is obviously superior to that of an existing imaging grading method. According to the present invention, the intervertebral disc degeneration plasma protein diagnosis system is established based on the SOMAscan technology for the first time, the protein concentration threshold of the ELISA verification is provided, the efficient complementation with the MRI Pfirrmann grading is achieved, the advantages of noninvasive property, objective property and high sensitivity are provided, and the new method is provided for the clinical early screening and dynamic monitoring.

The invention relates to the technical field of agricultural organic water-soluble fertilizers, in particular to an organic water-soluble fertilizer composition containing a rhizosphere enzyme-sensitive peptide nanofiber carrier and growth-promoting bacteria and application of the organic water-soluble fertilizer composition. The composition in the claim 1 is prepared from the following components in parts by mass: bifunctional oligopeptide which is self-assembled to form nanofibers, the surface layer of the bifunctional oligopeptide is grafted with rhizosphere signal peptide, and the middle layer and the core of the bifunctional oligopeptide contain amino acid-rich nutritional peptide fragments; filtering the dried growth promoting bacillus biocenosis; and auxiliary organic auxiliary materials. During use, protease secreted by a plant root system firstly cuts the surface layer of the nanofiber, and the signal peptide is released to activate the root system; then, the signal peptide promotes the flora to proliferate, the flora secretes more proteases, fiber cores are cut, nutrient peptide is slowly released, and the purposes that the root system is activated firstly and then continuous oxygen supply is achieved are achieved. The technology obviously enhances the absorption efficiency of root branches and nutrients, reduces the fertilizer loss, improves the crop yield and the stress resistance, and the carrier and the fungicide can be completely biodegraded, and are environment-friendly and safe.

The invention discloses a multiple fluorescent staining method for multiple protein targets, and relates to the technical field of multiple immunofluorescent staining. According to the method provided by the invention, the number of dyeable protein targets in the tissue slice can be greatly increased, multiple fluorescent staining steps can be simplified, and by adopting the method provided by the invention, imaging can be carried out after all the targets are directly stained on the basis of one tissue slice. The number of tissue slices required by dyeing imaging is reduced, dyeing of more than 6 protein targets can be completed by only one tissue slice, and errors caused by slice continuity differences are avoided. In addition, repeated elution and re-dyeing are not needed, so that the problems of antigen loss and weakening and the like caused by repeated elution are avoided.

The invention provides a method for separating multiple proteins in a milk raw material and application of milk protein.The method comprises the steps that the temperature of feed liquid is controlled to range from 50 DEG C to 55 DEG C, the influences of salt balance and beta-casein dissociation caused by the temperature are eliminated, and the stability of the milk protein in the feed liquid is kept; and then whey protein and casein are separated through chymosin treatment. Specifically, in the invention, chymosin treatment is carried out in a first reaction solution, so that whey in the first reaction solution can be separated out, which is caused by the fact that a k-CN micelle net-shaped structure is continuously rearranged in a curding process, and the whey distributed in the net-shaped structure is transferred. In the invention, by controlling the pH of a reaction system and the mass ratio of calcium to phosphorus in the chymosin treatment process, the content of whey separated out from the first reaction liquid can be remarkably increased, so that the yield of lactoprotein is further increased.

The present invention relates to a recombinant nodular dermatosis virus (LSDV) vector comprising an expression cassette encoding a foot and mouth disease virus (FMDV) polyprotein and a modified FMDV 3C protease. The invention also relates to a composition comprising the recombinant LSDV vector and an expression cassette. The invention further relates to a dual vaccine against LSDV and FMDV.