240 results about "Cell protein" patented technology

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 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 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 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 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 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 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.

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 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.

This invention relates to a protein feed prepared from biorefined baijiu lees and its preparation method, belonging to the fields of bioengineering and feed resource development technology. It includes the following steps: (1) subjecting the baijiu lees to acid pretreatment and dual-enzyme synergistic hydrolysis sequentially to obtain a hydrolysate containing fermentable sugars; (2) adding Candida utilis culture and urea for carbon-nitrogen synergistic liquid fermentation to obtain a Candida utilis viable count ≥4.0 × 10⁻⁶. 8 Fermentation broth with CFU / mL; drying is then performed. This invention pretreats baijiu (Chinese liquor) lees under high pressure with dilute acid, followed by synergistic hydrolysis with two enzymes, and then utilizes *Candida utilis* for carbon-nitrogen synergistic liquid fermentation, achieving efficient utilization of glucose and xylose, with a cell protein yield ≥0.70 g / L / h. This invention realizes the high-value utilization of all components of baijiu lees, providing a process route with industrial-scale production potential for the biorefining of baijiu lees to prepare protein feed.

The invention relates to the technical field of bioengineering, in particular to a preparation method and application of an antibacterial peptide, and the amino acid sequence of the antibacterial peptide is shown as SEQ ID No: 1 and is named as L8. The preparation method of the antibacterial peptide L8 comprises the following steps: S1, obtaining an L8 target gene segment; connecting an L8 target gene segment with a pLC vector to obtain a connection product; transforming the connection product into an escherichia coli DH5-alpha competent cell, and screening positive clones to obtain a pLC-L8 recombinant plasmid; s2, taking the pLC-L8 recombinant plasmid as a substrate, and carrying out cell-free protein synthesis reaction to obtain the antibacterial peptide L8, the cell-free protein synthesis system comprises 175 mM of potassium glutamate, 10 mM of ammonium glutamate, 2.7 mM of potassium oxalate, 33 mM of PEP, 38 mM of compound amino acid, 10 mM of magnesium glutamate, 2% (w / v) PEG-8000, 5 nM of pLC-L8 recombinant plasmid, 11.25 mM of lactose, 33.33% (v / v) of crude extract, 100 mg / mL of escherichia coli tRNA and 2.5 nM of T7 RNA polymerase. The antibacterial peptide L8 is obtained by optimizing the antibacterial peptide LRGG, and compared with the LRGG, the antibacterial peptide L8 subjected to cell-free protein synthesis reaction expression has better antibacterial activity and biological safety.

The invention provides a recombinant herpes simplex virus and a herpes simplex virus modification method. The herpes simplex virus modification method comprises the step of replacing an ICP4 protein coding gene in a herpes simplex virus genome containing an infected cell protein 4 (ICP4) gene with an hTERTp-fluorescent protein expression cassette. The expression cassette comprises an hTERTp promoter and a fluorescent protein coding sequence controlled by the hTERTp promoter, and the transcription direction is opposite to that of an ICP4 promoter in a genome. The hTERTp-fluorescent protein expression cassette in the recombinant virus obtained by the method disclosed by the invention can be normally expressed in response to telomerase activity. Therefore, the recombinant virus is capable of expressing a fluorescent protein, such as mBaoJin, in a cell having human telomerase activity. Cells infected by the virus can be identified through fluorescence signals, and the higher the telomerase activity is, the stronger the fluorescence intensity is. The virus has wide application value in research of tumor action mechanisms and stem cell action mechanisms, health assessment, screening of tumor drugs, research and development of diagnostic reagents and establishment of animal models.

The present invention discloses a Schizochytrium strain that efficiently converts food waste to produce single-cell protein and its application, belonging to the field of bioengineering technology. The Schizochytrium spLOX‑2 deposit number is: CGMCC No.41623. The present invention also discloses a method for efficiently converting food waste to produce single-cell protein, fermenting food waste to produce volatile fatty acids; and using it as a carbon source to culture Schizochytrium to obtain single-cell protein. The recombinant strain spLOX‑2 in the present invention has improved utilization of lactic acid, and its biomass and protein yields are increased by 14.19% and 12% respectively compared with the wild-type strain, and the oil content is significantly increased by 107.1%. The present invention provides a new path for a sustainable bioeconomic model, in which waste is converted into valuable biochemicals, laying a solid foundation for expanding the scale of the process and the economic feasibility in industrial applications.

The present invention pertains to the use of miRNA technology for improving recombinant production of polypeptides of interest in host cells. Expression cassettes are provided which produce a miRNA targeting and down-regulating a host cell protein which interferes with production of the polypeptide of interest.

The invention discloses a monoclonal antibody 5B11 for recognizing an extracellular domain of peste des petits ruminants virus H protein and application of the monoclonal antibody 5B11, and belongs to the technical field of monoclonal antibodies. According to the invention, PPRV-H-185 protein with an amino acid sequence as shown in SEQ ID NO.1 is used for immunizing a mouse, a monoclonal antibody 5B11 with a neutralizing effect is prepared, a heavy chain variable region of the monoclonal antibody 5B11 comprises CDR1-3 with an amino acid sequence as shown in SEQ ID NO.7-9, and a light chain variable region of the monoclonal antibody 5B11 comprises CDR1-3 with an amino acid sequence as shown in SEQ ID NO.12-14. Experimental results show that the monoclonal antibody can be combined with a virus, so that the virus loses the ability of combining an epithelial cell H protein receptor Nectin-4 and a lymphocyte H protein receptor SLAM, and the virus is prevented from invading cells. The invention provides a new biological material for detection and prevention of peste des petits ruminants virus, and has a good clinical application prospect.

The present disclosure provides materials and methods related to the purification of a biologic from a biological fluid. In particular, the present disclosure provides compositions, and related methods, comprising peptide ligands 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 biological fluids during the production of a biologic.

The invention belongs to the technical field of biology, and particularly relates to Meyerozyma caribba XX2120, and the preservation number of the Meyerozyma caribba XX2120 is GDMCC No: 67777. The invention further relates to a preparation method of the Meyerozyma caribba XX2120. The strain is obtained by taking saccharomycetes X2120 as a starting strain, carrying out ARTP mutagenesis treatment and screening, has relatively strong acid resistance, relatively high salt stress resistance and certain alcohol resistance and aldehyde resistance, can effectively utilize monosaccharide, disaccharide and trisaccharide, has a wide utilization range on carbohydrate and has a relatively good utilization rate on inorganic nitrogen. Moreover, the strain can be used for producing mycoprotein through liquid fermentation by taking Luzhou-flavor distillers' grains as a raw material, and also can be used for performing solid fermentation on the Luzhou-flavor distillers' grains, so that the true protein content of the distillers' grains is increased, the nutritional value of the distillers' grains is improved, and the recycling of distillers' grain resources is effectively promoted.

This invention discloses a method for detecting hepatocellular carcinoma (HCPs) in biological protein drugs based on LC-MS / MS. The method involves covalently binding HCP-derived cell line protein samples and monoclonal antibody proteins to the samples using chemical reagents, followed by mixing and fractionation. Finally, liquid chromatography-mass spectrometry (LC-MS / MS) analysis is performed to obtain the analytical results for HCPs in the monoclonal antibody drug. Compared with existing technologies, this invention enhances the detection signal by adding host cell proteins from monoclonal antibody production, significantly improving detection sensitivity and the depth of identification of low-abundance proteins. This fills a significant gap in HCP analysis and can be used to detect and demonstrate HCP clearance, particularly extremely low-level hydrolases in drug manufacturing processes, which has a crucial impact on ensuring the quality of biological protein pharmaceutical products and patient safety.

The application discloses a preparation method of glutamic acid bacterial protein ecological fertilizer, and the preparation steps are as follows: firstly, zinc polysilicate is added into glutamic acid fermentation waste liquid, and bacterial protein-zinc polysilicate complex is obtained through filtration; then, zinc silicate and hydroxyapatite are added, and amino acid composite hydroxyapatite particles are formed through the action of citric acid; and finally, urea-humic acid-dipotassium hydrogen phosphate complex is sprayed on the amino acid composite hydroxyapatite particles to obtain the ecological fertilizer. The ecological fertilizer can not only fully utilize the bacterial protein in the glutamic acid fermentation waste liquid, but also play a good slow-release effect of nitrogen, phosphorus and other elements through the action of two complexes. In addition, the addition of silicon and zinc elements can play a supplementing effect for the growth of crops, so that the fertilizer has the dual functions of environmental protection and long-acting slow release, and has a good promoting effect on the growth of crops.

The invention discloses candida utilis engineering bacteria and a method for producing single-cell protein by fermenting corn thin pulp, and belongs to the technical field of biology. According to the candida utilis engineering bacteria disclosed by the invention, the candida utilis is subjected to gene modification, a chitin synthase gene in the candida utilis is knocked out, and an exogenous keratinase gene is integrated, so that the candida utilis engineering bacteria can utilize corn thin pulp as a substrate for fermentation. In the fermentation process, carbon and nitrogen sources in the thin corn pulp can be effectively utilized for growth, macromolecular protein in the thin corn pulp can be hydrolyzed and converted into single-cell protein, the content of the single-cell protein in final fermentation liquor is increased, the protein content of the final fermentation liquor can reach 60% or above, and the proportion of the single-cell protein can reach 30% or above; the corn thin pulp can be efficiently utilized, and meanwhile, single-cell protein in the fermentation liquor can be used for adding animal feed so as to meet the requirement for animal protein feed in the market.

The present disclosure relates to methods, cells, and compositions for producing a product of interest, e.g., a recombinant protein. In particular, the present disclosure provides improved mammalian cells expressing the product of interest, where the cells (e.g., Chinese Hamster Ovary (CHO) cells) have reduced or eliminated activity, e.g., expression, of certain host cell proteins, e.g., enzymes including, but not limited to, certain lipases, esterases, and / or hydrolases.

The invention relates to the technical field of microbial fermentation, and provides a method for producing single-cell protein, which comprises the following steps: inoculating a crop straw matrix with a first microbial seed for first fermentation; the first microbial seed comprises trametes versicolor; sterilizing a fermentation product obtained by the first fermentation; inoculating second microbial seeds into the sterilized fermentation product for second fermentation; the second microbial seed comprises yarrowia lipolytica / candida utilis. According to the scheme, the combination of the selected first microbial seed solution and the second microbial seed solution has a remarkable synergistic effect, so that the crude protein content, the lignin removal rate, the cellulose retention rate and the yeast biomass are improved, and the hemicellulose loss rate is reduced.

The object of the present invention is to provide a protein production method capable of producing an active protein with high efficiency even at a low temperature, and a cell-free protein synthesis kit. A protein production method including producing a protein with a reaction solution of a cell-free protein synthesis system containing either one or both of a cold shock protein and a nucleic acid containing a coding region encoding an amino acid sequence of the cold shock protein. A cell-free protein synthesis kit including one or both of a cold shock protein and a nucleic acid containing a coding region encoding an amino acid sequence of the cold shock protein, and a reaction solution of a cell-free protein synthesis system.

The present invention relates to a novel fermentation, extraction and purification process for obtaining a single-cell protein for use in the food industry, using a particular strain of Kluyveromyces marxianus referred to as KMPROTEO 1, which results in a product consisting of a single-cell protein powder with a protein content equal to or greater than 60%, that is highly bioavailable, which, since it is soluble, can be included in different food matrices, without changing the formulation or the profile of the product to which it is added, which we have named "PRO60". The process disclosed in this specification includes the description of the aerobic fermentation of the strain of Kluyveromyces marxianus referred to as KMPROTEO 1, as well as the use of the ingredient or final product obtained, by adding same to some preferred food matrices.

The invention relates to the technical field of waste resource utilization, in particular to preparation of a biogas slurry nanofiltration permeate culture medium and a method for applying the biogas slurry nanofiltration permeate culture medium to continuous production of single-cell protein. The method comprises the following steps: carrying out matrix filtration on biogas slurry obtained by carrying out anaerobic fermentation on livestock and poultry manure, and carrying out membrane filtration on filtrate of the biogas slurry after the matrix filtration; the matrix filtration is enhanced by adopting a coagulant, the membrane filtration is carried out step by step by adopting ultrafiltration and nanofiltration, and the obtained permeate is subjected to C / N and pH regulation. The biogas slurry nanofiltration permeate culture medium provided by the invention is suitable for growth and metabolism of candida utilis, the protein synthesis efficiency is remarkably improved, the nanofiltration permeate is high in purity, interference of infectious microbes and impurities is reduced, the nutritional value and safety of a product are improved, and efficient production of single-cell protein and resource utilization of biogas slurry can be realized.

The present disclosure provides materials and methods related to the purification of viral vectors. In particular, the present disclosure provides compositions, and related methods, comprising peptide ligands 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 biological fluids during the production and purification of adeno-associated viruses (AAVs).