64 results about "Polyproteins" patented technology

The invention relates to the technical field of fluorescence immunoassay, and discloses a quantum resonance fluorescence immunoassay medical detection method, which comprises the following steps: 1, carrying out fixation and permeation treatment on cells, respectively incubating the cells by using labeled different quantum dot-antibody compounds, specifically binding antibodies with corresponding target proteins in the cells, and detecting the target proteins in the cells; a labeled cell sample is obtained; 2, under a confocal microscope, laser with a specific wavelength is adopted to excite the marked cell sample, and fluorescence lifetime image data of different quantum dot emission wavelengths are collected; according to the method, the fluorescence lifetime image data of different quantum dot emission wavelengths are acquired through the confocal microscope, the position and content information of different target proteins can be clearly distinguished, simultaneous analysis and accurate positioning of multiple target proteins are realized, and an efficient means is provided for multi-protein research in a complex biological system.

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.

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.

The invention discloses a plasma protein-based stomach cancer early-stage multi-molecular diagnosis model construction method, and aims to solve the problems that the existing stomach cancer early-stage diagnosis standard and minimally invasive or non-invasive method are imperfect, the sensitivity and specificity of the existing diagnosis molecular marker are low and the like, an Olink proteomics PEA technology is utilized and a specific detection panel is selected to construct a stomach cancer early-stage multi-molecular diagnosis model. And identifying the protein markers of gastric cancer, early gastric cancer and gastric precancerous lesions in a discovery queue (n = 88). In a discovery queue, a group of multi-protein molecular marker models consisting of 13 proteins are successfully identified by utilizing an LASSO regression model, and the multi-protein molecular marker models have good diagnostic performance for distinguishing gastric cancer, early gastric cancer and pregastric lesion and contrast in the discovery queue and verification pair. Furthermore, in a verification queue sample, the diagnosis model also shows good diagnosis performance, and the marker model is expected to become a powerful tool for early diagnosis of gastric cancer.

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 invention relates to the technical field of biological medicine detection, and particularly discloses an electrochemical luminescence kit for rapidly detecting tumor markers, which comprises a capture molecule, an electrochemical luminescence ECL labeled molecule, a biotin labeled detection antibody, a streptavidin fixed plate and a liquid reagent, according to the present invention, the capture molecule, the ECL labeling molecule, the biotin labeling detection antibody and the streptavidin fixation platform interact to form the novel multi-protein complex, such that each target antigen can initiate multiple signal conversion during the reaction process; theoretically, the tetramolecular structure can obviously improve the signal transmission efficiency, so that the low-abundance tumor marker can be quickly and accurately detected, and the high-specificity recognition of the target tumor marker is realized by capturing molecules and detecting antibodies. The high affinity between biotin and streptavidin is utilized, so that non-specific binding is greatly reduced, background signals are reduced, and the accuracy and reliability of detection are improved.

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.

The present disclosure provides a method for increasing a number of proteins identified, including: S1. adding a binding buffer and an aluminosilicate zeolite to a sample to be tested to obtain a first suspension; S2. incubating the first suspension, subjecting an incubated first suspension to first high-speed centrifugation to obtain a first supernatant and a first precipitate, discarding the first supernatant, and retaining the first precipitate; S3. repeating a process as follows several times: resuspending the first precipitate with a washing buffer to obtain a second suspension, subjecting the second suspension to second high-speed centrifugation to obtain a second supernatant and a second precipitate, discarding the second supernatant, and retaining the second precipitate; and S4. preparing the second precipitate into a protein sample, and subjecting the protein sample to mass spectrometry detection.

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 disclosure provides heterodimeric polypeptides comprising: 1) a variant hepatitis C virus (HCV) E2 polypeptide and an HCV E1 polypeptide; 2) a variant HCV E1 polypeptide and an HCV E2 polypeptide; or 3) a variant HCV E1 polypeptide and a variant HCV E2 polypeptide, where the variant HCV E2 polypeptide and / or the HCV E1 polypeptide comprises one or more T cell epitopes, present in an HCV polypeptide other than an HCV E1 polypeptide or an HCV E2 polypeptide. The present disclosure provides nucleic acids encoding a polyprotein that includes E1 and variant E2, E2 and variant E1, or variant E2 and variant E1. The present disclosure provides a method of producing an E1 / E2 heterodimer of the present disclosure. The present disclosure provides a method of inducing an immune response in an individual. The present disclosure provides variant E2 polypeptides and variant E1 polypeptides; and nucleic acids encoding same.

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.