22 results about "INSULIN HUMAN" patented technology

The invention discloses an interchangeable active human insulin solution standard substance as well as a preparation method and application thereof, and belongs to the technical field of stoichiometric analysis and detection. The preparation method provided by the invention comprises the steps of purification of human insulin, self-assembly of the human insulin, confirmation of an insulin self-assembly compound structure, purification of active human insulin, determination of immunoaffinity activity concentration and specific activity, and preparation of the active human insulin solution standard substance with interchangeability. The preparation method provided by the invention can be used for calibration of a human insulin conventional in-vitro diagnosis system, development of an in-vitro diagnosis detection reagent, method performance verification and value traceability of a human insulin detection result in serum, so that the accuracy and comparability of a clinical detection result are ensured, and public health and life safety are protected.

The present invention relates to a test kit for performing an assay to detect and measure at least one autoantibody molecule in a subject developing type 1 diabetes (T1D). The invention more specifically relates to a recombinant antigen molecule fused to a bioluminescent reporter protein to detect an autoantibody molecule against a self-antigen. Said self-antigen is preferably expressed by pancreatic islet beta cells and include insulin, proinsulin, Glutamate decarboxylase 65, islet antigen 2, and Zinc transporter 8. More specifically, the present invention relates to an assay comprising a set of at least three recombinant antigen molecules in a single composition, the antigen molecules being selected from the list comprising recombinant human Glutamate decarboxylase 65, recombinant human islet antigen 2, recombinant human Zinc transporter 8, recombinant human insulin, and recombinant human proinsulin. The invention further relates to methods for determining the presence and / or level of at least one autoantibody molecule in a sample from a human subject developing T1D using said recombinant antigen molecules. According to the invention, the presence, absence, or amount of one or more autoantibody molecules against said self-antigens is determined with a luciferase immune precipitation system (LIPS) assay or a modified version of a LIPS assay, i.e., a solid phase capture LIPS (scLIPS) assay.

This invention belongs to the field of immune cell culture technology, specifically relating to an in vitro expansion culture medium for NK cells and its culture method. It consists of a basal culture medium and auxiliary additives. The basal culture medium is X-VIVO 15 medium; the auxiliary additives include the following components: human serum albumin (HSA), IL-12, IL-18, IL-15, rapamycin, nicotinamide, gelatin-IL-15 microspheres, alginate-IL-21 microspheres, recombinant human insulin, transferrin, glutamine, β-mercaptoethanol, a mixture of non-essential amino acids and lipids. This invention achieves time-dependent synergistic IL-15 sustained-release / IL-21 pulsed release and metabolic synergistic NAD+ release. + Mitochondrial / lipid membrane homeostasis enables efficient amplification and long-lasting cytotoxicity.

Provided is a method whereby it is possible to quickly and inexpensively quantify human insulin secreted from human cells. Specifically disclosed is a method for quantifying the insulin secretion ability of human cells, which is characterized by culturing a recombinant cell in which a gene expressing a fusion C-peptide and an insulin gene in a human cultured cell strain genome are replaced, and measuring the amount of the secreted fusion C-peptide. The fusion C-peptide is a fusion C-peptide obtained by inserting a detectable tag peptide into a C-peptide chain in a human proinsulin molecule.

The invention belongs to the technical field of cells, and particularly relates to a neural stem cell culture medium and a culture method. The neural stem cell culture medium comprises a basic culture medium and the following components added into the basic culture medium in terms of final concentration: 0.5-10 mg / L of human insulin, 20-30 [mu] g / L of epidermal growth factors, 2-3 [mu] g / L of folic acid, 2-5 [mu] g / L of N-glutamine, 10-40 [mu] g / L of N-acetylcysteine, 0.1-5 mg / L of ethanolamine, 0.5-2 mg / L of carnitine, 0.5-5 mg / L of vitexin glucoside and 15-25 [mu] g / L of plecanatide. The vitexin glucoside and the plecanatide play a synergistic effect, so that the rapid division and proliferation of the neural stem cells are remarkably promoted, the cell differentiation potential of the neural stem cells is maintained, and the clinical application of the neural stem cells can be fully guaranteed.

The application discloses a kind of active human insulin solution standard substance with interchangeability and its preparation method and application, belong to chemical metrology analytical detection technical field.The preparation method provided in the application includes the purification of human insulin, self-assembly of human insulin, confirmation of insulin self-assembly complex structure, the purification of active human insulin, determination of immunological affinity active concentration and specific activity, preparation of active human insulin solution standard substance with interchangeability.The above preparation method provided in the application can be used for the calibration of human insulin routine in vitro diagnostic system, the development of in vitro diagnostic detection reagent and method performance verification, and for the value traceability of human insulin detection result in serum, ensure the accuracy and comparability of clinical detection result, protect public health and life safety.

The application relates to the technical field of stem cells, in particular to a mesenchymal stem cell culture system without heterogeneity and application thereof. The mesenchymal stem cell culture system without heterogeneity comprises a basic culture medium and an additive component, wherein the basic culture medium is DMEM / F12, and the additive component comprises: recombinant human transferrin, recombinant human epidermal growth factor, recombinant human basic fibroblast growth factor, recombinant human insulin, recombinant human interferon gamma, hydrocortisone, vitamin A, vitamin D2, vitamin B, human blood albumin, CTS KnockOut SR XenoFree, glutamine and dextran. The mesenchymal stem cell culture system without heterogeneity provided by the application improves the proliferation efficiency of primary culture and in-vitro expansion culture of mesenchymal stem cells, is high in safety, good in stability and suitable for large-scale popularization and application.

This invention relates to a kit and method for detecting anti-human insulin-like growth factor binding protein 7 (IGFBP-7) using a double-antibody sandwich principle combined with a nucleic acid probe self-assembly signal amplification method. The kit comprises three parts: 1) a capture domain formed by IGFBP-7 antibody 1 coated on the surface of an ELISA plate; 2) a tandem functional domain composed of biotin-labeled IGFBP-7 antibody 2, streptavidin, and biotin-labeled DNA primers; and 3) a fluorescence signal functional domain composed of two DNA hairpin probes. Compared with traditional ELISA methods, the method of this invention significantly improves detection sensitivity and features high specificity, high sensitivity, good stability, and strong versatility, as well as good clinical application value.

The invention provides a product for treating type 2 diabetes mellitus based on a stem cell insulin gene vector. The product consists of mesenchymal stem cells modified by genetic engineering and a cell preservation solution, the genetic engineering modified mesenchymal stem cells are obtained by transducing human mesenchymal stem cells by using a recombinant lentiviral vector, and the recombinant lentiviral vector can express a fusion protein composed of three parts of human proinsulin, a linker peptide and a GLP-1 receptor agonist. According to the application, human proinsulin and a GLP-1 receptor agonist are constructed into a single fusion protein, human mesenchymal stem cells are transduced by utilizing a lentiviral vector, and then a standardized three-stage induced differentiation process is carried out, so that a cell therapy product which can intelligently respond to blood glucose changes for a long time and has stable functions is finally obtained. The defects of transient treatment effect, single secretion function and large cell product batch-to-batch difference in the prior art are fundamentally overcome.

The present invention relates to the technical field of cell culture, and specifically relates to a mesenchymal stem cell culture and amplification medium and a preparation method thereof. The mesenchymal stem cell culture and amplification medium uses DMEM / F12 medium as a basic component, and the DMEM / F12 medium further includes the following components: oxygen-releasing material 8 g / L, natural extract 2 g / L, growth factor 30 ng / mL, nicotinamide 5 mM, cholesterol 8 μg / mL, recombinant human transferrin 20 mg / L, recombinant human insulin 10 mg / L, vitamin C 4 mg / L, L-glutamine 2 mM, reduced glutathione 60 μg / L, sodium selenite 20 mg / L. The prepared mesenchymal stem cell culture and amplification medium is beneficial to the growth and proliferation of mesenchymal stem cells.

Provided are a novel insulin derivative and a use thereof. Comparing the novel insulin derivative with human insulin, by carrying out amino acid mutation on sites B17, B25, and other sites such as the A14 site, the A19 site, the B3 site, and the B4 site, the novel insulin derivative has good performance of resisting in-vivo protease degradation; by means of an acylation reaction, a fatty acid side chain is connected at the B29 site, so that the fatty acid side chain can reversibly bind to albumin, further increasing the presence duration in the body, and thereby achieving a long-acting action mechanism. The novel insulin derivative shows obvious action characteristics of long-acting insulin, and can be used for developing a human insulin analogue administered once a week.

The invention relates to the field of pharmaceutical analysis, in particular to a method for determining the content of phenol and m-cresol in an insulin preparation based on GC-MS / MS (Gas Chromatography-Mass Spectrometry / Mass Spectrometry). The method comprises the following steps: (1) precisely measuring human insulin injection or insulin glargine injection, putting into a measuring flask, diluting by 100 times with a methanol aqueous solution, uniformly shaking, precisely measuring, putting into the measuring flask, diluting by 10 times with the methanol aqueous solution, and taking as a test solution; (2) a quartz capillary column is adopted, the column temperature is kept at 200 DEG C for 5 min, the sample size is 1 microliter, the split ratio is 10: 1, and carrier gas is inert gas; (3) an ionization source is an electron bombardment ion source, the energy is 70 eV, the temperature of the ion source is 200 DEG C, the interface temperature is 250 DEG C, the solvent delay is 1 min, the mass scanning range is 45-500 m / z, the electron multiplication voltage is 3.0 kV, the collision gas is inert gas, and multi-reaction monitoring MRM and full scanning Scan are collected at the same time. The specificity, the linear range, the accuracy, the repeatability, the stability and the durability of the method are superior to those of the prior art.

The invention relates to a pharmaceutical composition containing human insulin analogue fusion protein and application thereof. The pharmaceutical composition has good biological activity and stability.

The invention discloses a human umbilical cord mesenchymal stem cell culture medium and a preparation method thereof, and the human umbilical cord mesenchymal stem cell culture medium comprises a basal culture medium, a growth factor, an antioxidant, recombinant human insulin, carotenoid, poly (adenosine diphosphate ribose polymerase) and catechin. The concentration of the recombinant human insulin is 1mg / L to 5mg / L, the concentration of the carotenoid is 10mu g / mL to 100mu g / mL, the concentration of the poly adenosine diphosphate ribose polymerase is 1mu M to 100mu M, and the concentration of the catechin is 10mu g / mL to 100mu g / mL. According to the present invention, with the human umbilical cord mesenchymal stem cell culture medium and the preparation method thereof, the rapid in-vitro proliferation of the human umbilical cord mesenchymal stem cells can be easily achieved, and the cell replicative aging can be improved.

The present invention relates to a nucleic acid molecule comprising a nucleotide sequence encoding a metabolic protein or a portion thereof, or a sequence having at least 90% sequence identity to said metabolic protein or a portion thereof, a human insulin-like growth factor II (IGFII) gene sequence, and a nucleotide sequence encoding at least one peptide that promotes cellular uptake or transcytosis inserted into said IGFII gene sequence between the nucleotides encoding amino acids 28 and 42 of mature IGFII. The invention further relates to related viral particles, fusion proteins, and uses thereof.

The present application relates to the field of animal cell culture medium, and specifically discloses a serum-free culture medium for SP2 / 0 cell culture and a preparation method and application. The serum-free culture medium for SP2 / 0 cell culture is calculated per 1L, and the raw materials used for its preparation include 10.15g of MEM-α dry powder containing nucleosides, 50-1000μg heparin sodium, 200-4000mg recombinant human serum albumin and corresponding amounts of arachidonic acid, tocopherol acetate, Pluronic F-68, transferrin, recombinant human insulin, sodium selenite, ethanolamine, ascorbic acid, hydrocortisone, etc., and the remainder is water for cell culture. The serum-free culture medium for SP2 / 0 cell culture of the present application can meet the requirements that SP2 / 0 cells can be stably suspended and cultured under serum-free conditions without taming. During the SP2 / 0 cell culture process, when reaching high density, the cells do not aggregate, and the multiplication is stable.

The invention relates to nucleic acid molecules comprising a nucleotide sequence encoding a metabolic protein or a part thereof or a sequence having at least 90% sequence identity to said metabolic protein or part thereof, a human insulin-like growth factor II (IGFII) gene sequence, and a nucleotide sequence encoding at least one peptide that facilitates cellular uptake or transcytosis which is inserted at a location between the nucleotides encoding amino acids 28 and 42 of mature IGFII of said IGFII gene sequence. The invention further relates to related viral particles, fusion proteins and uses thereof.

The invention discloses an induction method of dopaminergic neural stem cells, and relates to the technical field of cell culture and neurobiology, and the induction method comprises the following steps: inoculating neural stem cells into an induction culture medium, culturing under the conditions of 37 DEG C and 5% CO2, and carrying out induced differentiation for 6-14 days to obtain dopaminergic neurons; the induction culture medium takes DMEM / F12 as a basic culture medium, and contains an effective amount of the following components: human insulin, progesterone, hydrocortisone, dexamethasone, estradiol, transferrin, an epidermal growth factor (EGF) and a fibroblast growth factor (FGF). The induction culture medium and the method have the advantages that the survival of neural stem cells can be effectively supported; and the gene is guided to directionally differentiate towards the dopaminergic neuron lineage. The method is clear in operation flow and stable in differentiation process, the expression of the characteristic marker of the dopaminergic neuron can be detected in the obtained cell population, and a stable and reliable cell source is provided for related scientific research.

This invention relates to the field of NK cell culture medium technology, and discloses an NK cell culture medium, its preparation method, and its applications, including: an activation phase culture medium, an expansion phase culture medium, and a maturation phase culture medium; wherein the three culture media include a common combination of recombinant human transferrin, recombinant human insulin, polyvinyl alcohol, ethanolamine, ascorbate-2-phosphate, α-thioglycerol, sodium selenite, ferric citrate monohydrate, and nicotinamide, as well as different combinations of cytokines. This invention uses polyvinyl alcohol instead of recombinant human serum albumin. Polyvinyl alcohol can form a physical protective layer to stabilize the cell membrane and act as a surfactant to disperse hydrophobic nutrients, thereby more efficiently and stably supporting cell survival and proliferation. The culture medium of this invention, combined with the three-stage culture method of this invention, can effectively maintain high cytotoxicity of cells and significantly reduce the expression of functional exhaustion-related molecules while obtaining high-purity NK cell expansion.

A human insulin-like growth factor (IGF) binding protein stock solution and method of making the same include a non-recombinant human IGF binding protein-3 (nr-IGFBP-3) in an aqueous buffered medium. The concentration of the nr-IGFBP-3 in the stock solution ranges from about 16 micrograms per milliliter (μg / ml) to about 40 μg / ml. A kit includes a set of calibrators for nr-IGFBP-3. The set of calibrators includes the nr-IGFBP-3 in different concentrations.

The invention discloses a culture medium for promoting separation of Wharton's jelly mesenchymal stem cells, application and a method, and belongs to the technical field of mesenchymal stem cell culture. The culture medium comprises a basic culture medium, a recombinant human basic fibroblast growth factor, a recombinant human hepatocyte growth factor, recombinant human insulin, recombinant human transferrin, a recombinant human stromal cell-derived factor (SDF-1alpha), lycopene and a ginseng extract. The culture medium provided by the invention is free of serum and heterologous components, the climbing-out speed and number of Wharton's jelly mesenchymal stem cells can be remarkably increased, and the dryness of the mesenchymal stem cells can be well maintained.