313 results about "Cell culture" patented technology

This invention discloses a monoclonal antibody against human hepatitis B e antigen and its application, relating to the field of hepatitis B detection technology. In the heavy chain variable region of the monoclonal antibody, the amino acid sequences of the complementarity-determining regions (CDR1-3) are as shown in SEQ ID NO. 1-3 or have at least 95% homology with the sequences shown in SEQ ID NO. 1-3; in the light chain variable region, the amino acid sequences of the complementarity-determining regions (CDR1 and CDR3) are as shown in SEQ ID NO. 4 and 5 or have at least 95% homology with the sequences shown in SEQ ID NO. 4 and 5, and the amino acid sequence of the complementarity-determining region (CDR2) is FAS. The monoclonal antibody provided by this invention can effectively recognize natural hepatitis B e antigen, and the titer of the supernatant from B cell culture can reach 1:10000, making it fully applicable to the detection and research of human hepatitis B e antigen. The chemiluminescent reagent prepared using the monoclonal antibody described in this invention has advantages such as high specificity, strong anti-interference ability, high detection sensitivity, and good stability, with almost no missed detections, and can replace imported reagents.

The invention relates to the technical field of synthetic biology, metabolic engineering and cell culture. The present invention provides a cell for the production of a negatively charged, preferably sialylated, oligosaccharide wherein the cell is genetically engineered to have or express, preferably overexpress, a hydrolytic UDP-N-acetyl-D-glucosamine-2-epimerase. The invention further provides the use of said cells in culture or incubation. Also described are methods of producing negatively charged, preferably sialylated oligosaccharides using the cells, and purification of the negatively charged, preferably sialylated oligosaccharides.

The present invention relates to a cell biomass (e.g. food product or pet food product) or its components prepared from cultivated metazoan cells, method of its preparation and a system for its preparation.

The invention provides bovine ephemeral fever virus positive serum as well as a preparation method and application thereof. The preparation method comprises the following steps: culturing bovine ephemeral fever virus by using MDBK suspension cells, inactivating by using beta-propiolactone, concentrating by using a 500kd hollow fiber column to obtain an antigen solution, and emulsifying the antigen solution and 61VG adjuvant according to a mass ratio of 1: 1.5 to prepare the vaccine; carrying out primary immunization and secondary immunization on the receptor, collecting blood of which the serum neutralizing antibody titer is greater than 1: 4096 14-21 days after the secondary immunization, and separating to obtain positive serum. The positive serum can be used for specific test, identification test and the like of the bovine ephemeral fever virus, can also be used for preparing related reagents, kits and standard substances, and provides efficient and normative technical support for prevention and control of bovine ephemeral fever.

The invention provides 3D tumor cell culture hydrogel as well as a preparation method and application thereof. The 3D tumor cell culture hydrogel is prepared from the following raw materials: polyvinylpyrrolidone and polyvinyl alcohol. The preparation method comprises the following steps: mixing a polyvinylpyrrolidone solution with a polyvinyl alcohol solution, adjusting the pH value, and freezing-unfreezing to obtain the 3D tumor cell culture hydrogel. The 3D tumor cell culture hydrogel provided by the invention does not need to be additionally added with a chemical cross-linking agent, and the material is high in biocompatibility and flexible in modulus adjustment, and can be well applied to a 3D in-vitro culture model of tumor cells.

The invention provides a culture method of avian encephalomyelitis virus, and belongs to the technical field of biology. According to the method, chicken lymphoma suspension cells are adopted to culture the avian encephalomyelitis virus, the titer of the obtained avian encephalomyelitis virus reaches 106.5-6.8 EID50 / 0.2 mL, and the method is short in culture period, high in production efficiency, low in cost, controllable in quality and capable of being used for large-scale industrial production.

The invention discloses a sleep-promoting nasal spray based on mesenchymal stem cell-derived exosomes and preparation and application thereof.The preparation method comprises the steps that 1, mesenchymal stem cells separated from umbilical cord tissue are cultured to P3-P5 generations, cell culture supernate is collected, cells are digested, and then cell debris and macromolecular impure protein are removed through pretreatment; (2) filtering and concentrating the pretreated cell culture supernatant and replacing a part of solution; (3) carrying out degerming filtration on the solution obtained in the step (2) through a 0.22 [mu] m filter membrane to obtain an MSC-Exos solution; and (4) adding a stabilizer with a proper concentration into the MSC-Exos solution to obtain a compound MSC-Exos solution, namely the sleep-promoting nasal spray. The product is a cell-derived natural active substance, can significantly improve sleep, and does not involve any pharmaceutical ingredient.

The invention discloses a preparation method of a cochlear organ chip integrated with a blood labyrinth barrier, and belongs to the field of biomedical engineering. The preparation method of the simulated blood labyrinth barrier comprises the following steps: constructing an endothelial cell culture cavity and a pericyte culture cavity, and separating the endothelial cell culture cavity and the pericyte culture cavity through a simulated basement membrane; and respectively inoculating percutaneous cells and endothelial cells into a percutaneous cell culture cavity and an endothelial cell culture cavity, and culturing to obtain the simulated blood lost barrier. The chip can effectively simulate the structure and the function of the BLB, and the barrier integrity of the BLB is evaluated through TEER (trans-epithelial electrical resistance) measurement and an apparent permeability coefficient. The invention also relates to application of the chip in NIHL drug screening, in particular to drug evaluation for oxidative stress induced inner ear organ injury. Through the platform, the protection effect of the candidate drugs on the TBHP-induced oxidative stress injury of the inner ear organs can be evaluated, and a new tool and thought are provided for developing novel NIHL treatment drugs.

The invention discloses a bioreactor multi-parameter adaptive control system and method based on machine learning, and belongs to the technical field of bioreactor intelligent control, and the bioreactor multi-parameter adaptive control system comprises a bioreactor hepatocyte culture unit, a data acquisition and control unit and a machine learning intelligent decision-making unit. And the data acquisition and control lower computer controls operation variables such as gas supply, sampling and perfusion in the culture unit execution mechanism, collects culture process data returned by the parameter detection module, constructs a data-driven cell state prediction model by utilizing machine learning, and optimizes model parameters and operation variables based on a prediction result to realize closed-loop control. The method can overcome the defect that there is no precise mechanism model for hepatic cell culture, and multi-parameter collaborative optimization can be realized only depending on process data, so that the state of the hepatic cells is sensed in real time, full-cycle optimization of cell viability and functions is realized, and the activity and density of hepatic cell culture are effectively improved.

The invention discloses a culture method for improving proliferation and differentiation capacity of NK (Natural Killer) cells based on angelica sinensis-astragalus membranaceus exosomes, which comprises the following steps: firstly extracting exosomes in angelica sinensis and astragalus membranaceus, then adding the exosomes into an NK cell culture system according to a specific concentration, and regulating and controlling an NK cell signal channel by virtue of active ingredients carried by the exosomes so as to improve the proliferation and differentiation capacity of the NK cells. Therefore, the proliferation rate and differentiation maturity of the NK cells are remarkably improved. Experimental results show that compared with a traditional culture method, the proliferation multiple of the NK cells cultured through the method is increased by 28.99%-33.69%, the proportion of differentiated mature cells is increased by 15.91%-28.30%, and the cell killing activity is not remarkably reduced. The method can be widely applied to the field of immune cell treatment, provides a high-quality NK cell source for clinic, effectively solves the problems of slow cell proliferation rate, low differentiation maturity, easy activity reduction after large-scale culture and the like in traditional NK cell in-vitro culture, and has important clinical application value and industrialization prospect.

The invention provides a gamma delta T cell culture method and a culture medium, and belongs to the technical field of T cell culture. Modified nanoparticles, zoledronic acid, vitamin C and glutamine are added into a 1640 culture solution, the concentration of the modified nanoparticles ranges from 0.5 g / L to 1 g / L, the concentration of the zoledronic acid ranges from 0.5 mu mol / L to 1.5 mu mol / L, the concentration of the vitamin C ranges from 200 mu mol / L to 500 mu mol / L, and the concentration of the glutamine ranges from 0.1 mmol / L to 0.2 mmol / L. The prepared culture medium can efficiently stimulate amplification of the gamma delta T cells and improve the killing activity of the gamma delta T cells, meanwhile, the survival rate of the gamma delta T cells is also improved, and meanwhile, the culture method of the gamma delta T cells is simple, low in cost and wide in application prospect.

The invention provides a pharmaceutical composition for treating non-small cell lung cancer and application of the pharmaceutical composition. The pharmaceutical composition comprises gefitinib, erlotinib, afatinib, neratinib, osimertinib and the like which serve as active ingredients. The pharmaceutical composition disclosed by the invention can simultaneously contain various EGFR inhibitors as active ingredients, and the active ingredients are controlled at ultra-low dosage, so that the pharmaceutical composition not only can ensure the treatment effect, even the drug effect of the pharmaceutical composition is obviously superior to that of a high-dosage EGFR inhibitor independent administration group, but also can ensure the safety, and the pharmaceutical composition is suitable for clinical application. And after the active components are combined, a good synergistic effect is achieved, and more importantly, the occurrence of drug resistance can be remarkably reduced or even resisted. In a lung cancer cell culture experiment, the EGFR inhibitor independent administration group has a drug resistance phenomenon at the earliest 20 days, but the pharmaceutical composition group still has no drug resistance phenomenon after 160 days of administration.

The invention provides a stem cell culture process information abnormity early warning method, which comprises the following steps: preprocessing original data in a stem cell culture process, extracting layered features to obtain basic statistical features, time sequence trend features and environmental factor interaction features, and updating the features in real time through a dynamic sliding window, after the verified feature set is obtained, fusion calculation is carried out to obtain a multi-dimensional fusion feature set; calculating a trend deviation degree, a feature correlation degree and a fluctuation dispersion degree of the multi-dimensional fusion feature set, and carrying out dynamic weight distribution and fusion to obtain a fusion anomaly degree value; and matching the fusion anomaly degree value with a multi-level threshold value to obtain a matching result, judging an abnormal state type in combination with an association relationship between the feature anomaly type and the cultivation stage, and generating an anomaly early warning result. According to the method, through data preprocessing, layered feature extraction and fusion and dynamic anomaly calculation, the accuracy, real-time performance and practicability of stem cell culture anomaly early warning are improved.

The invention discloses a skin-tightening anti-wrinkle essence and a preparation process thereof, and relates to the technical field of cosmetic preparation, and the preparation process comprises the following steps: obtaining animal umbilical cord tissue, culturing, collecting cell culture supernatant after culture, carrying out low-speed centrifugation to remove cells and dead cells, and carrying out concentration, filtration and sterilization through a tangential flow membrane technology to obtain the skin-tightening anti-wrinkle essence. The animal umbilical cord extract is obtained; the preparation method comprises the following steps: mixing the animal umbilical cord extract with water, butanediol, glycerol, hydroxypropyl tetrahydropyrantriol, 1, 2-hexanediol, trehalose, p-hydroxyacetophenone, allantoin, hydrolyzed sodium hyaluronate, sodium hyaluronate, tetrahydromethylpyrimidine carboxylic acid, xanthan gum and essence, homogenizing under a stirring condition, and standing for defoaming. By constructing an active stable microenvironment and an efficient transdermal delivery system, the technical problem that the animal umbilical cord extract serving as a core component is easy to inactivate and difficult to absorb is solved, and a faster and more stable skin structure remodeling effect is realized by utilizing the synergistic effect of the animal umbilical cord extract and functional components.

This invention provides a lung stem cell peptide for treating pulmonary nodules, its preparation method, and its application, belonging to the field of biopharmaceutical technology. The invention involves inducing iPSCs to differentiate into lung stem cells; culturing the lung stem cells, centrifuging, collecting the supernatant to obtain crude lung stem cell solution; mixing the crude lung stem cell solution with a protease for enzymatic hydrolysis, centrifuging, concentrating, and spray-drying to obtain the lung stem cell peptide for treating pulmonary nodules. This invention optimizes the composition of each culture medium to induce iPSCs to differentiate into lung stem cells, then extracts and separates lung stem cell peptides from these cells. The extracted lung stem cell peptides are intravenously reinfused into the lungs, effectively inhibiting the formation of pulmonary tumor nodules, significantly reducing the number of pulmonary tumor nodules, and without toxic side effects or adverse reactions.

The present invention relates to a method for producing farnesene oil, the method comprising the use of a raw material fermentation yeast in a fermentation vessel to produce a whole cell culture fluid (WCB); and adding a surfactant to the fermentation vessel to reduce emulsification within the fermentation tank.

The invention discloses a purification method of cord blood mesenchymal stem cell source exosome, and relates to the technical field of bioengineering. The method comprises the following steps: firstly, carrying out pH regulation pretreatment on cell culture supernate through a technical path of combining gradient centrifugation and membrane filtration, and sequentially removing cell debris and large-particle impurities by adopting multi-stage differential centrifugation; then carrying out selective protein precipitation by utilizing polyethylene glycol with specific molecular weight, and realizing fine separation of the exosome by combining sucrose-meglumine diatrizoate density gradient centrifugation; and finally, terminal purification is completed under low-temperature and low-pressure conditions through a polyethersulfone membrane filtration system with an optimized pore size. According to the method, centrifugal parameters, reagent concentration and filtering conditions are precisely regulated and controlled, so that the particle size of the obtained exosome is intensively distributed in the range of 30-150nm, the protein purity reaches 95% or above, the recovery rate exceeds 85%, and the separation efficiency and quality of the exosome are remarkably improved.

The utility model relates to the technical field of stem cell culture, and discloses efficient stem cell sampling equipment which comprises a sampling box, the top of an assembly cavity is fixedly connected with a mounting plate, a sampling barrel is embedded in the center of the mounting plate, a rotating mechanism is arranged in the center of the sampling barrel, and the rotating mechanism is connected with the mounting plate. The top of the rotating mechanism is connected with a containing mechanism, the containing mechanism comprises a containing disc, clamping grooves, a first sampling assembly, a mounting groove and a second sampling assembly, the multiple clamping grooves are formed in the circumference of the edge of the containing disc at equal intervals, the first sampling assembly is connected into the clamping grooves in a clamped mode, and the second sampling assembly is connected into the mounting groove in a clamped mode. According to the equipment, the accommodating mechanism is arranged, the sampling assembly I and the sampling assembly II are used for stably placing the sampling tube and the sampling box on the accommodating disc, and the accommodating disc is driven by the rotating mechanism to rotate so as to realize sampling, so that the adaptability of the equipment to different sampling containers is improved, and the equipment is convenient to operate and replace parts.

The present invention relates to improved platelet-derived growth factor (PDGF) polypeptide variants, particularly polypeptides comprising the sequence of any one of SEQ ID NOs: 2 to 4 or a sequence having at least 80% similarity thereto, wherein the PDGF polypeptide exhibits increased stability, biological activity and / or solubility compared to the wild-type PDGF polypeptide of SEQ ID NO: 1. The present invention further provides nucleic acid sequences encoding the PDGF polypeptides of the present invention, and also provides a method for growing animal cells, comprising culturing the animal cells in a culture medium containing the PDGF polypeptide of the present invention.

The invention discloses an efficient purification process of a bispecific antibody and a special chromatography system thereof. The efficient purification process of the bispecific antibody comprises the following steps: pretreatment: performing 0.22 mu m microfiltration on a cell culture supernatant to remove cell debris, and performing deep filtration to reduce turbidity to be less than or equal to 1NTU so as to reduce subsequent chromatography medium pollution; carrying out improved Protein A affinity chromatography, namely carrying out affinity chromatography by adopting an agarose medium modified by a recombinant Protein A ligand, taking 20mM Tris-HCl as a loading buffer solution, and taking the pH value of the loading buffer solution as 7.4, taking 100mM citric acid as an elution buffer solution, and taking the pH value of the elution buffer solution as 3.2-3.5; performing mixed mode chromatography: adopting a composite mode medium containing a benzamidine functional group, taking 50mM sodium phosphate as a buffer solution, adjusting the pH value to 6.0-6.5, adjusting the NaCl concentration to 0.1-0.3 M, and specifically removing a double-antibody polymer and a mismatched isomer by utilizing a hydrophobic effect and an ion exchange synergistic effect; and performing nanoscale filtration and terminal treatment: removing viruses by adopting a virus filtration membrane with the aperture of 30nm, and performing 0.22 mu m sterile filtration to obtain a final product. According to the process, the purification efficiency, the yield and the product purity are remarkably improved, and the process is suitable for efficient purification of the bispecific antibody.

The utility model relates to the field of culture solution concentration detection, in particular to an on-line protein concentration detection device for expanding a concentration range. The device comprises a first injection pump, a selector valve, a container assembly, a dilution and detection assembly and an air filter, the selector valve is provided with a public port and six selection ports uniformly distributed around the public port, and the public port is communicated with one of the six selection ports; a buffer ring is communicated between the first injection pump and the common port of the selection valve; the container assembly comprises a calibration standard liquid container, a first cleaning liquid container, a second cleaning liquid container and a culture container; the dilution and detection assembly comprises a three-way valve, a dilution assembly and a detection assembly respectively communicated with two ends of the three-way valve, and a sample reserving valve communicated with the output end of the detection assembly. According to the utility model, the detection concentration range is expanded to 6.5 orders of magnitude, the requirement of the protein measurement concentration range from extremely low concentration to extremely high concentration in the whole cell culture process is met, and the measurement accuracy and sensitivity are high.

According to the stem cell culture solution pollution-free collection module, the sterile isolation and vacuumizing process of the internal environment of a collection bottle is achieved through a collection assembly, the function that air in the bottle is pumped out firstly and then the stem cell culture solution is injected is achieved through the coaxial inner needle and outer needle design of an infusion assembly, and the stem cell culture solution is collected through a multi-stage self-sealing structure after the needles are pulled out. And the collection bottle is automatically closed and sealed after liquid injection is completed, so that the pollution-free stem cell culture solution collection module which is pollution-free in the whole process and is closed and safe is realized. The device is characterized in that the device comprises a collecting assembly and an infusion assembly, the infusion assembly is inserted into the collecting assembly, the infusion assembly injects a stem cell culture solution into the collecting assembly, and the collecting assembly comprises a collecting bottle, a sealing plug, a disposable sterile unsealing film, a duckbill valve, a check valve and a multi-layer self-resetting diaphragm; and the sealing plug is clamped at the opening end of the collecting bottle.

The invention discloses a biological cell culture sampling device for a biological test, which comprises a culture bottle and a sealing cover made of a transparent material, a rotary locking mechanism and a sealing assembly are arranged between the sealing cover and a sampling port in a matched manner, and an electric push rod is fixedly arranged at the upper end of the sealing cover. The piston end of the electric push rod extends into the sealing cover and is fixedly provided with a sampling needle assembly, the side wall of the sealing cover is communicated with a vacuumizing connector and a pressure regulating air inlet valve, an efficient particle air filtering film is arranged in the pressure regulating air inlet valve, and the top of the sealing cover is provided with a suction connector used for being connected with external negative pressure suction equipment. Through cooperation of the rotary locking mechanism and the sealing assembly, reliable sealing of the sealing cover and the sampling opening of the culture bottle is achieved, vacuumizing pretreatment is combined, external air is prevented from entering the culture bottle during puncture sampling, the sterility of a culture system is guaranteed, a main sealing structure of the culture bottle does not need to be opened in the whole sampling process, and the sampling efficiency is improved. And the original stable gas environment in the culture bottle is maintained.

1. Name of the design product: Controller for controlling small cell culture tank. 2. Use of the design product: for controlling oxygen flow, stirring speed, acid and base addition ratio (i.e. pH value) and temperature of small cell culture tank. 3. Design points of the design product: in shape. 4. Picture or photo best indicating the design points: front view.

The invention relates to the technical field of biological medicine, and discloses an anti-human 5T4 single-chain antibody, the antibody has the capability of specifically binding to human 5T4 protein, the human 5T4 protein corresponds to TPBG / WAIF1, and the GenBank login number is NP001018111.1; the antibody also has the capability of specifically binding and expressing human 5T4 protein tumor cells, and the human 5T4 protein tumor cells are breast cancer cells MCF-7, prostate cancer cells PC3 or gastric cancer cells MGC-803. By combining single B cell culture with a magnetic bead sorting technology, natural pairing of a heavy chain and a light chain of an antibody can be reserved, and the defect that pairing is damaged by a traditional hybridoma technology is overcome; meanwhile, false positive clones are effectively eliminated through double positive screening of detecting protein binding activity through ELISA and detecting cell binding activity through FACS, the finally obtained targeting 5T4 monoclonal antibody can be specifically combined with human 5T4 protein and tumor cells expressing the protein, and a high-quality targeting molecular basis is provided for research and development of subsequent antibody drugs and CAR-T cell treatment products.

The application discloses a kind of stem cell culture devices, belong to stem cell technical field, including sealed incubator, syringe, the inside of the sealed incubator is equipped with electric rotating table, the top of the electric rotating table is equipped with adjustable shaking assembly, the adjustable shaking assembly is equipped with petri dish carrier, petri dish limiting component, the petri dish limiting component is equipped with uniform feeding assembly, the top of the sealed incubator is equipped with liquid injection pipe, the top of the liquid injection pipe is equipped with sealed iris component, the outside of the sealed incubator is equipped with lifting assembly.The adjustable shaking assembly, petri dish limiting component, uniform feeding assembly of being equipped with, so that petri dish realizes multidimensional periodicity shaking, promotes stem cell to adhere to the growth and material exchange;Sealed iris component, syringe are set, in the process of injecting reagent, the sealing of incubator is always maintained, avoid exposing sample in air, guarantee the stability of culture environment.

The invention relates to the technical field of virus detection, in particular to a fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection probe 1106VP and / or 1106WP and primers 1106VWF and 1106VWR. The invention also discloses a fluorescent RT-PCR detection reagent, a fluorescent RT-PCR detection kit, a fluorescent RT-PCR detection probe, a primer, the fluorescent RT-PCR detection reagent and the application of the fluorescent RT-PCR detection kit in identification of a porcine reproductive and respiratory syndrome NJ-1106R strain and a porcine reproductive and respiratory syndrome non-NJ-1106R strain. The probe and the primer disclosed by the invention are high in specificity and high in sensitivity; the accuracy is high, and the accuracy rate of blind sample detection is 100%; the kit is high in universality, can be used for detecting virus liquid for cell culture in a laboratory, and can also be used for detecting porcine serum and tissue samples in clinical samples. The method is an effective method for identifying the PRRSV NJ-1106R vaccine strain and other PRRSV wild strains.

This invention provides methods for culturing animal cells and related technologies. [Solution] Animal cells are cultured in the presence of lecithin.

The present invention generally pertains to methods of cultivating cells and producing a recombinant protein of interest, such as dupilumab. In particular, the present invention pertains to the use of a fixed seed schedule to determine the expansion time of seed train unit operations for improved cell growth and protein titer. The protein produced by the fixed seed schedule, such as dupilumab, can be later subjected to lipase activity reduction methods.

The application provides a factor composition for activating DC cells and a method for application thereof, and belongs to the technical field of cell preparations.The factor composition for activating DC cells provided by the application comprises the following components: rhGM-CSF, rhIL-4, rhSCF, rhTNF-alpha, IFN-alpha, insulin and lycium barbarum polysaccharide.Through the synergistic effect between the components, the application can not only promote the differentiation of PBMC into DC cells, increase the proportion of DC cells, increase the initial amount of DC cells, promote the proliferation of DC cells, increase the activity of DC cells, but also can increase the expression of antigen costimulatory molecules and reduce the culture cycle of DC cells.