404 results about "Cell protein" patented technology

The invention belongs to the technical field of biology, and particularly relates to kluyveromyces marxianus with high beta-glucan content and application of the kluyveromyces marxianus. Specifically, the invention provides a Kluyveromyces marxianus, the classification name of the Kluyveromyces marxianus is Kluyveromyces marxianus CJA0020, and the preservation number of the Kluyveromyces marxianus is CCTCC (China Center For Type Culture Collection) No.M 20241673. According to the Kluyveromyces marxianus screened by the invention, the strain growth speed is high, a strain fermentation product has higher biomass and beta-glucan content, and the beta-glucan content can reach 1.23 + / -0.06 g / L; the biomass can reach 400 g / Kg (wet weight) and the protein content can reach 45% when the strain is cultured for 32 hours during fed-batch fermentation, and the strain can also be used for producing yeast single-cell protein.

The invention discloses a single-cell protein extraction and hydrolysis process. The invention discloses a single-cell protein extraction and hydrolysis process which comprises the following steps: S1, adding a chemical reagent into single-cell protein thalli to lyse cells to obtain a cell lysis solution; s2, adding a protein denaturation reagent into the cell lysis solution, and centrifuging to obtain a protein denaturation precipitate; s3, adding a protein dissolving reagent into the protein denaturation precipitate to obtain a protein solution; and S4, adding enzyme into the protein solution, and carrying out protein hydrolysis to obtain a hydrolysate. The method has a wide application prospect, and is expected to promote technical progress and industrial development in related fields.

The invention belongs to the technical field of biological detection, and discloses a kit and a method for quantitatively detecting residual quantity of host cell protein of pichia pastoris in order to solve the problems of limit of quantitation and low detection sensitivity of an existing detection method. According to the present invention, the empty plasmid without the recombinant human serum albumin gene is introduced into the pichia pastoris, and purification is performed by combining the Protein A chromatographic column and the host cell protein coupling affinity column, such that the prepared polyclonal antibody can be effectively used for the quantitative detection of the residual amount of the pichia pastoris host cell protein in the biological product; the kit formed by the polyclonal antibody has good accuracy and repeatability on the quantitative detection of the host cell protein residues in the pichia pastoris source biological products, the quantification limit is 0.2 ng / mL, and the kit provided by the invention can meet the quantitative detection of the host cell protein residues in all the pichia pastoris source biological products.

The invention relates to the technical field of medicine analysis and detection, and particularly discloses a method for detecting the content of protein in a 3D-MLA adjuvant, which comprises the following steps: (1) preparing a test solution: firstly dissolving the 3D-MLA adjuvant by using a hydrochloric acid solution, performing 300-600W ultrasonic treatment for 2-5min, centrifuging 10000-13000g for 4-7min, and taking supernate; adding a compound enzyme into the supernate for enzymolysis; then adding concentrated hydrochloric acid, uniformly mixing, hydrolyzing for 20 + / -2 hours in a nitrogen atmosphere at 105-115 DEG C, blow-drying, and dissolving with a hydrochloric acid solution to obtain a test sample stock solution; adding a derivatization reagent into the test sample stock solution, and carrying out derivatization treatment, extraction and dilution to obtain a test sample solution; and (2) high performance liquid chromatography determination. According to the detection method provided by the invention, the residual quantity of the host cell protein in the 3D-MLA can be rapidly and accurately determined, and the quality control of the 3D-MLA is realized.

The invention discloses a method for detecting HCPs in a biological protein drug based on an LC-MS / MS method. The method comprises the following steps: covalently binding an HCPs-derived cell line protein sample and a monoclonal antibody protein with the sample by adopting a chemical reagent, mixing, fractionating, and finally performing liquid-phase mass spectrometry to obtain an analysis result of HCPs in the monoclonal antibody drug. Compared with the prior art, the detection signal is enhanced by adding the host cell protein for producing the monoclonal antibody, the detection sensitivity and the identification depth of low-abundance protein are remarkably improved, the important blank of HCPs analysis is filled up, and the monoclonal antibody can be used for detecting and proving HCPs removal, especially extremely-low-level hydrolase in drug process development, and can be applied to detection and proving of HCPs removal. The method has important influence on ensuring the quality of biological protein pharmaceutical products and the safety of patients.

The invention provides a pichia pastoris strain for efficiently converting methanol to produce high-methionine-content single-cell protein and application of the pichia pastoris strain, and belongs to the technical field of biology. According to the invention, a strain of pichia pastoris (named as Pichiapastoris CN-SCP 00 with a preservation number of CCTCC (China Center For Type Culture Collection) NO: M2025657) is obtained through separation, and is subjected to multiple rounds of ARTP iterative mutation breeding, so that a strain Pichiapastoris CN-SCPYE421 (with a preservation number of CCTCC NO: M2025658) for efficiently converting methanol to produce single-cell protein containing high methionine is obtained. Compared with a commercial strain, the Pichiapastoris CN-SCPYE421 has higher methanol tolerance and utilization capacity, the fermentation time is remarkably shortened, and in addition, the Pichiapastoris CN-SCPYE421 can well grow at the temperature of 35 DEG C. The Pichiapastoris CN-SCPYE421 is subjected to fermentation culture in a 5L fermentation tank, the dry weight of thalli reaches 120-150g / L, the protein content reaches 65-75% of the dry weight of the thalli, the Pichiapastoris CN-SCPYE421 is rich in various essential amino acids, and the methionine content reaches 2.3%. The invention provides an excellent strain for industrial development of single-cell protein, and has great social significance and economic value for the feed protein industry.

The invention relates to a Kluyveromyces marxianus strain with high protein yield and application of the Kluyveromyces marxianus strain, which is characterized in that the Kluyveromyces marxianus strain is firstly cultured on a slant, then is transferred into a seed culture medium for seed preparation, and then is inoculated into a fermentation tank, and yeast protein is obtained by adopting index feeding; the bacterial strain is a dominant mutant strain Kluyveromyces marxianus YWX-1, the protein content in a dry thallus reaches 55% or above and is increased by 26% or above compared with that of an original bacterial strain, the bacterial strain can be used for production of single-cell protein, the essential amino acid index (EAAI) value in the protein reaches 1.2, and the bacterial strain is rich in arginine, threonine and lysine and can be applied to production of single-cell protein. These amino acids play a key role in immune regulation, intestinal mucosal barrier function and muscle protein synthesis of young animals. The YWX-1 kluyveromyces marxianus strain has a wide application prospect in the production of neoplasm protein.

The invention provides a cell protein degradation platform based on an artificial biological macromolecular aggregate. Specifically, the invention provides a PROTAC functional module based on an interworking nucleic acid skeleton, a PROTAC-aggregate complex (MLO-PROTAC), a kit, application and a preparation method of the PROTAC functional module, the PROTAC-aggregate complex (MLO-PROTAC) and the kit, and also provides a targeted protein degradation method. In a PROTAC platform built by the PROTAC functional module and the PROTAC-aggregate complex, a programmable nucleic acid component is used as a core assembly unit, and the functional module is enriched and spatiotemporal-spatial regulation is performed by using the polypeptide aggregate, so that the delivery efficiency and the cytoplasm exposure degree are remarkably improved while the universality is maintained, and the delivery efficiency and the cytoplasm exposure degree are remarkably improved. Therefore, a more effective target protein degradation way is provided for the field.

The invention relates to a cell lysis solution, which is prepared from the following active components: 0.08 to 0.2 percent of DDM (Dichloromethane), 35 to 45 mM of CAA (Ceamic Acid Acetic Acid), 5 to 15 mM of TCEP (Trichloroethylene Phosphate), 85 to 100 mM of TEAB (Tetraethyl Ammonium Bromide), 0 to 0.25 percent of Protease MAX and 0 to 0.25 percent of RapiGest SF (SF). The invention further provides a single-cell protein pretreatment method based on the digital microfluidic system, the cell lysis solution is used for lysis, the number of detectable proteins and the number of detectable peptide fragments can be high, and the method can be well used for single-cell proteomics analysis.

Recombinant proteins that stimulate an immune response in the presence of naturally inhibitory ligand binding are described. The recombinant proteins include an extracellular domain of an inhibitory immune cell protein and an intracellular domain of a stimulatory immune cell protein connected via a transmembrane domain. The recombinant proteins can be used to stimulate immune cell activation in the fight against cancers and infectious diseases, among other uses.

The application provides a multi-gas control device, a use method and an application, and relates to the technical field of biological gas fermentation. The device comprises a gas supply module, the gas supply module comprises multiple groups of gas sources and connecting pipelines, each gas source is connected to an air inlet pipeline through a respective connecting pipeline, a secondary pressure reducing valve, a T-shaped filter, a mass flow meter, a stop valve, a regulating valve and a check valve are sequentially arranged on the connecting pipeline, the air inlet pipeline is connected to a liquid circulation pipeline, gas is discharged from a spraying mechanism connected to the liquid circulation pipeline, the spraying mechanism is arranged in a fermentation tank, the spraying mechanism is connected to a circulating pump through the liquid circulation pipeline, the circulating pump is connected to the bottom of the fermentation tank through a pipeline, the top of the fermentation tank is connected to a detection pipeline, and a precision filter, a vacuum pump, a drying device, a pre-analyzer and a gas analyzer are sequentially arranged on the detection pipeline. The application realizes efficient absorption and utilization of multiple gases, and simultaneously obtains high yield and high quality single-cell protein.

The invention discloses a double-carbon-source mixed fermentation metabolism regulation system for single-cell protein production, and belongs to the technical field of fermentation engineering. The method solves the problems that in the prior art, the independent utilization efficiency of methanol and methane is low, the metabolic pathways of microorganisms to the two carbon sources are different, and high-efficiency single-cell protein production is difficult to realize, and by comprehensively analyzing various data related to historical parameters in the historical parameters, the production efficiency of the single-cell protein is improved. According to the method, key metabolic nodes having significant influence on the yield and quality of the single-cell protein can be accurately identified, and reasonable weights are distributed for the key metabolic nodes, so that the production quality of the single-cell protein in fermentation engineering is optimized, and the realization of a production target is ensured; meanwhile, according to the production target of the actual fermentation engineering, the optimization cost of the key metabolism nodes is evaluated, excessive optimization of high-cost nodes is avoided, the economic benefit of single-cell protein production is improved, the cost budget of the fermentation engineering is reduced, and the flexibility of the fermentation engineering and the scientificity of metabolism regulation decision are enhanced.

The invention provides a bispecific antibody and a purification method thereof. The purification method comprises the following steps: a) loading a sample containing a bispecific antibody to a affinity chromatographic column, and leaching and eluting; the biomacromolecules in the sample are adsorbed on the affinity chromatography column in the leaching stage, the biomacromolecules comprise antibodies, host cell proteins and protein aggregates, and the antibodies comprise bispecific antibodies and homodimers; separating the antibody from the affinity chromatography column in the elution stage, and collecting the eluate in the elution stage; b) loading the eluent to a hydrophobic membrane material, carrying out flow-through, and collecting a flow-through liquid containing the purified antibody; and the flow-through liquid used for flow-through contains 0.73 to 0.77 M of (NH4) 2SO4. The method can solve the problem that homodimers in bispecific antibodies are difficult to efficiently remove in the prior art, and is suitable for the field of antibody purification.

The present disclosure provides T cell modulatory polypeptide-epitope conjugates (T-Cell-MP-epitope conjugates) comprising a chemical conjugation site at which an NY-ESO (e.g., NY-ESO-1 or NY-ESO-2) or a MAGE (e.g., MAGEA4) peptide epitope is covalently attached and at least one immunomodulatory polypeptide sequence that may be variant selected to exhibit reduced binding affinity to its cognate co-immunomodulatory polypeptide. The T-Cell-MP-epitope conjugates are useful for modulating the activity (e.g., increasing proliferation or cytotoxic activity) of T-cells specific to the NY-ESO or MAGE peptide epitope in an epitope selective / specific manner, and accordingly, for treating individuals with, for example, cancers expressing the covalently attached epitope. The disclosure also provides T-Cell-MP-epitope conjugates with targeting sequences that can server to, among other things, localize the T-Cell-MP-epitope conjugates to a specific tissue or cell type.

The present disclosure relates to proteomics, including the detection of immune cell target proteins as examples that the disclosed methods can be used for protein detection. Methods, compositions, and systems are described for identifying the presence of proteins, including detecting immune cell proteins from a sample using a combination of multiplex immuno-PCR with antibodies conjugated to oligonucleotides comprising an antibody-specific barcode flanked by PCR priming sites; detection of barcoded amplicons obtained from immuno-PCR with encoded padlock probes comprising codes specific for the barcodes / target protein; rolling circle amplification (RCA) of the circularized padlock probes; and determination of the sequence of the codes amplified by RCA by next generation sequencing or using fluorescently labeled hybridization probes.

The invention provides a fermentation method for producing single-cell protein from corn straw. The fermentation method comprises the following steps: (1) inoculating a microbial seed solution into a crushed corn straw solid culture medium; (2) after the solid-state fermentation is completed, adding water, and reacting at 65-85 DEG C for 4-8 hours; and (3) cooling the liquid culture medium, and then inoculating a microbial seed solution for fermentation. By adopting the method, the yield of single-cell protein can be up to 142.68 g / L, and the yield of crude protein can be up to 44.8%. Compared with direct fermentation of the corn straw, the yield of single-cell protein and the content of crude protein can be remarkably increased, and the fermentation method for achieving efficient utilization of the corn straw is provided.

The invention discloses kluyveromyces marxianus and application of the kluyveromyces marxianus in production of single-cell protein by taking straw as a raw material. The Kluyveromyces marxianus is a ZH-2 bacterial strain which is CGMCC (China General Microbiological Culture Collection Center) No.32295 The strain is obtained from a wild strain in a laboratory adaptive evolution mode, wheat straw and an inorganic nitrogen source can be used as raw materials for efficiently producing single-cell protein, compared with a parent strain, the utilization speed and the utilization rate of biomass, glucose and xylose are remarkably increased, the yield of ZH-2 SCP can reach 0.047 g.g <-1 >, and the yield of ZH-2 SCP can reach 0.045 g.g <-1 >. And the essential amino acid mode basically accords with adult and infant amino acid modes, and can be used as high-quality protein nutrition supply. The Kluyveromyces marxianus ZH-2 strain capable of obtaining single-cell protein by fermenting wheat straw as a raw material is provided from the perspective of utilizing cheap waste resources, so that the production cost of SCP is greatly reduced, and the problem of protein resource shortage is effectively solved.

The invention discloses Candida tropicalis, and the preservation number of the Candida tropicalis in the Guangdong Microbial Culture Collection Center is GDMCC No: 64869. The Candida tropicalis has the advantages that the preservation number of the Candida tropicalis is CGMCC No: 64869; the invention further provides application of the candida tropicalis in fermentation production of single-cell protein, a culture medium suitable for the candida tropicalis and a method for fermentation production of the single-cell protein. The candida tropicalis is a high-stress-resistance strain of vinegar residue hydrolysate, is suitable for large-scale production, can effectively promote recycling of vinegar residue waste biomass resources, and relieves the current situation of protein feed shortage in the breeding industry to a certain extent.

The invention provides a post-treatment method of single-cell protein fermentation liquor and a fermentation method of single-cell protein thalli. Nuclease and cell wall lytic enzyme are respectively added into fermentation liquor containing single-cell protein thalli to treat the thalli, so that nucleic acid and cell wall residues can be reduced, and the problems that in a traditional fermentation process, the application of the thalli in food is limited due to the high nucleic acid content of the thalli, and the protein extraction efficiency is reduced due to the existence of the cell walls are solved. In addition, by optimizing fermentation conditions, the protein content of the single-cell protein is remarkably increased. The process is simple, low in cost, suitable for industrial production and capable of being widely applied to the fields of feed, food and medicine.

The invention provides a methylobacterium strain JC03 for producing single-cell protein and application of the methylobacterium strain JC03 in the field of aquaculture, and belongs to the technical field of microorganisms. The methylobacterium strain JC03 is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.32616. The strain is high in growth efficiency, high in thallus crude protein content, rich in amino acid spectrum, high in essential amino acid content and rich in functional components for improving the immunocompetence of aquatic animals; the single-cell protein produced by the strain can completely or partially replace fish meal protein in feeds of marine fishes such as turbot, pseudosciaena crocea, grouper and rainbow trout; vegetable protein in feed of freshwater fish such as grass carp can be completely replaced; animal and plant proteins in feeds for shrimps and crabs, such as penaeus vannamei, can be completely replaced, and animal and plant proteins in stichopus japonicus feeds can be completely replaced; the immunocompetence of fed aquatic animals can be obviously improved. The invention provides a microbial strain for efficiently converting methanol to produce single-cell protein, and the microbial strain has a wide application prospect in the field of aquaculture.

The present disclosure provides materials and methods related to the purification of viral vectors. In particular, the present disclosure provides peptides, compositions, adsorbents, and related methods, capable of removing process-related impurities (e.g., host cell proteins, nucleic acids, and media components) and product-related impurities (e.g., product fragments, product aggregates, and inactive forms derived from product degradation by or association with other species in the cell culture harvest) from samples during the production and purification of adenovirus.

The invention relates to the technical field of feed mixing devices, in particular to a mixing device for Leymus chinensis single-cell protein feed processing, which comprises a stirrer, a stirring mechanism mounted on the stirrer, a dust removal mechanism, a spraying mechanism and a discharging mechanism, a premixing mechanism is mounted on the side surface of the stirrer, and a transmission mechanism is mounted on the dust removal mechanism; when forage is added into the stirrer and in the earlier stage of stirring, large feed dust can be raised, an axial flow fan is connected with an external power supply to suck out the feed dust on the stirrer, the feed dust is collected by a filter screen, clean air is discharged from the tail end of a connecting shell, and a first motor is started to drive a stirring shaft to rotate; the stirring shaft can drive the filter screen to rotate on the connecting shell after rotating, the filter screen can abut against the inner bottom of the connecting shell when rotating anticlockwise, the collected feed dust falls into the connecting hopper along with rotation of the filter screen, collection and treatment of the feed dust are facilitated, and air pollution and feed waste are avoided.

The invention relates to a saccharomyces lipolytica strain, a breeding method and application. The Yarrowia lipolytica strain is preserved in the China Center for Type Culture Collection (CCTCC) on July 22, 2024, and the preservation number of the Yarrowia lipolytica strain is CCTCC NO: M 20241641. When the arrowia lipolytica strain is used for preparing the single-cell protein, the conversion rate of the single-cell protein reaches 40% or above, a continuous fermentation process is adopted, the production efficiency is improved by 50% or above, and the production efficiency reaches 2.5 g / L / h or above.

The present invention relates to a method for producing recombinant proteins, in which a larger cellular morphology is produced by, for example, degrading tags (SsrA16 and SsrA21) by SsrA, regulating morphology-related genes (including but not limited to mreBCD, ftsABEIQWXYZ, sulA, minCDE), thereby enabling more intracellular proteins to be accommodated, thereby improving the production of recombinant proteins (such as MTG and Vip) and single cell proteins.

To provide methods of producing and purifying AAV viral vectors having low empty capsids, low host cell proteins, and / or low contaminating DNA, while retaining high potency; and to provide a pharmaceutical composition containing the vector.SOLUTION: A method of manufacturing an AAV viral vector comprises: a. culturing adherent cells; b. transfecting the adherent cells with one or more plasmids to enable production of the AAV viral vector; c. lysing the adherent cells to isolate the AAV viral vector; d. acidifying and clarifying the cell lysate of c; e. purifying the product of d using cation exchange chromatography; f. filtering the product of e using tangential flow filtration; g. ultracentrifuging the product of f in a cesium chloride buffer; h. collecting the AAV viral vector from the product of g; and i. formulating and filling the AAV viral vector collected in h. to produce a final pharmaceutical product.SELECTED DRAWING: None

The current invention reports a method for purifying an antibody by reducing the content of a host cell protein. The method employs a wash step with a low conductivity aqueous solution in an affinity chromatography.

The invention discloses an Escherichia coli culture medium and application thereof in preparation of cell extracts required by cell-free protein synthesis, the culture medium comprises an initial culture medium or an initial culture medium and a supplementary culture medium, the initial culture medium comprises fructose, lactose, magnesium sulfate, ammonium sulfate, dipotassium phosphate, monopotassium phosphate, citric acid, trace elements, a vitamin solution and a nitrogen source; the fed-batch culture medium comprises a carbon source and a nitrogen source, the carbon source is fructose and a magnesium sulfate solution, and the nitrogen source comprises a nitrogen source solution added with yeast powder and yeast peptone. According to the culture medium disclosed by the invention, the escherichia coli can maintain logarithmic phase growth and reach relatively high thallus density (OD600 is greater than or equal to 60), and the prepared escherichia coli cell extract is used for cell-free protein synthesis, has high activity consistent with that of a cell extract obtained by low-density fermentation, and can be used for expressing various foreign proteins. According to the invention, the problem of low thallus density during preparation of the cell-free protein synthetic cell extract is solved, and the expression yield of the cell-free protein is effectively improved.

The invention discloses a separation method of single-cell protein fermentation liquor. The separation method comprises the following steps: 1) adding a filter aid into fermentation liquor containing single-cell protein thalli to form mixed liquor; 2) filtering the mixed solution through a plate frame to form a filter cake; 3) introducing nitrogen to purge the filter cake to remove moisture in the filter cake; and 4) carrying out flash evaporation drying on the dewatered filter cake to obtain a single-cell protein product. According to the process, filter cakes are formed through plate and frame filtering and squeezing, and then single-cell protein thalli are obtained through a flash evaporation drying technology. The process not only improves the extraction efficiency of the single-cell protein, but also reduces the production cost, and is suitable for large-scale production.

The disclosure relates to methods of detecting Host Cell Proteins (HCPs) in biopharmaceutical samples containing a drug product. More specifically, the disclosure relates to methods of assaying HCPs in biopharmaceutical samples using enzymatic digestion protocols, liquid chromatography (LC) gradients, immunoaffinity capture protocols, and High-Resolution Mass Spectrometry (HRMS) to enhance the recovery, detection, and profiling of HCPs, which can impact the efficacy, shelf-life, and therapeutic response of drug products.