66 results about "INSULIN HUMAN" patented technology

Disclosed and claimed is a new insect cell line, Sf900+, ATCC CRL-12579. The insect cell line was established from Lepidoptera, Noctuidae, Spodoptera frugiperda Sf-9 (ATCC CRL-1711) through multiple rounds of limiting dilution and selection in a serum-free insect medium supplemented with added human insulin. The insect cell line is useful in BEVS or as an adjuvant and has many characteristics and advantages. Also disclosed and claimed are recombinant proteins from recombinant baculovirus expression in insect cells such as Sf900+ cells, for instance, HA, NA, EPO, CD4, CEA, and thrombospondin.

The present invention includes fully human, neutralizing, monoclonal antibodies against human Insulin-like Growth Factor Receptor-I (IGFR1). The antibodies are useful for treating or preventing cancer in a subject. Also included are methods of using and producing the antibodies of the invention.

The present invention relates to novel antibodies capable of binding specifically to the human insulin-like growth factor I receptor IGF-IR and / or capable of specifically inhibiting the tyrosine kinase activity of said IGF-IR receptor, especially monoclonal antibodies of murine, chimeric and humanized origin, as well as the amino acid and nucleic acid sequences coding for these antibodies. The invention likewise comprises the use of these antibodies as a medicament for the prophylactic and / or therapeutic treatment of cancers overexpressing IGF-IR or any pathology connected with the overexpression of said receptor as well as in processes or kits for diagnosis of illnesses connected with the overexpression of the IGF-IR receptor. The invention finally comprises products and / or compositions comprising such antibodies in combination with anti-EGFR antibodies and / or compounds and / or anti-cancer agents or agents conjugated with toxins and their use for the prevention and / or the treatment of certain cancers.

The invention discloses a serum-free culture medium for mesenchymal stem cells. The serum-free culture medium for the mesenchymal stem cells comprises the following ingredients: fibronectin at the final concentration of 25mu g / ml, basic fibroblast growth factors at the final concentration of 10ng / ml, human epidermal growth factors at the final concentration of 15ng / ml, recombinant human insulin at the final concentration of 1mg / ml, human transferrin at the final concentration of 0.55mg / ml, human blood albumin in a volume ratio of 5 percent, sodium selenite at the final concentration of 0.67mug / ml, L-carnitine at the final concentration of 5mM and resveratrol at the final concentration of 30mu M. When the mesenchymal stem cells are cultured by the serum-free culture medium for the mesenchymal stem cells, animal-derived serum is not contained, so infection risks can be controlled; the L-carnitine and the resveratrol which are added into the serum-free culture medium for the mesenchymalstem cells can effectively improve the growth state of the mesenchymal stem cells; and the growth speed of the mesenchymal stem cells is remarkably improved, and the biological characteristics of themesenchymal stem cells are kept unchanged.

Provided herein are methods and compositions for treating a subject suffering from a deficiency in α-L-Iduronidase in the CNS. The methods include systemic administration of a bifunctional fusion antibody comprising an antibody to a human insulin receptor and an α-L-Iduronidase. A therapeutically effective systemic dose is based on the specific CNS uptake characteristics of human insulin receptor antibody-α-L-Iduronidase fusion antibodies as described herein.

The invention relates to a neural stem cells medium and a method for performing human neural stem cells in-vitro long-term culture and amplification by using the neural stem cells medium. The neural stem cells medium comprises the following ingredients by weight proportion: 100-1000 micrograms of heparin sodium, 10-100 micrograms of vitamin E, 5-50 milligrams of insulin human recombinant, 0.5-5 milligrams of putrescine, 2-10 micrograms of sodium selenite, 2-10 milligrams of human transferrin, 2-10 micrograms of progestin, 300 milligrams of L-glutamine, 5.9 grams of 2-[4-(2-Hydroxyethyl)-1-piperazine]ethanesulfonic acid, 10-100 micrograms of recombinant human epidermal growth factors, 10-100 micrograms of recombinant human basic fibroblast growth factors, 20-200 milligrams of vitamin C glucoside and 40,000-400,000 IU (international unit) of gentamicin. By the neural stem cells medium, the technical problems that human neural stem cells are easy to differentiate when cultured in vitro and long-term culture and amplification are difficult to implement are solved.

A connecting polypeptide has SEQ ID NO:73. A proinsulin polypeptide includes a mature insulin A-chain, a mature insulin B-chain, and a connecting peptide comprising SEQ ID NO: 73 linking the mature A-chain and the mature B-chain, wherein the connecting peptide is not a native human proinsulin C-peptide. The proinsulin polypeptides according to the invention can be made in high titers and in high purity.

The invention belongs to the technical field of stem cells and relates to a serum-free medium for placenta-derived mesenchymal stem cells and a preparation method thereof. The serum-free medium comprises a DMEM basic medium and further comprises vitamin H, glutathione, recombinant human insulin, human serum albumin, transferrin, fibroblast growth factors, epidermal growth factors, stem cell growth factors, Human stem cell factors and magnolol. The serum-free medium provided by the invention has the advantages of high safety and capability for significant improvement of proliferative capability of placenta-derived mesenchymal stem cells, the serum-free medium is capable of well keeping cell forms and stem cell characteristics of the placenta-derived mesenchymal stem cells and is convenient to popularize and apply.

The invention provides the immunomodulator with the free adjuvant having the function of preventing and curing the insulin-dependent diabetes mellitus. Aiming at the weak immunogenicity, the antigen epitope polypeptide gene (a length of antigen epitope p277 rooted from human HSP60) is associated repeatedly six times and inserted repeatedly into the backward position of the heat shock protein HSP65 gene rooted from the mycobacterium bovis, the fusing expression between the repeat series connecting antigen polyeptide and the HSP65 is realized. The produced fusing albumen did not need the adjuvant; the antigen polyeptide don't need be coupled with the carrier chemistry to the immunity. The fusing albumen can inspire the organism to produce the high titer aiming at p277 by means of the mucous membrane immunity, so the nosogeny of NOD chmice diabetic are depressed highly. The invention can prevent the 1 type and 1.5 type diabete characterized in depending on the insulin.

The invention relates to the technical field of cell culture, in particular to a culture medium, application thereof and a culture method. The culture medium comprises transferrin, recombinant human insulin, progesterone, fetal calf serum, stem cell growth factors, vitamin C, interleukin 2, interleukin 4, zinc sulfate and a DMEM / F12 culture medium. When the culture medium is used for culturing CD19-CAR-T cells, the cell proliferation multiple can be improved, the number of cell culture days is increased, and the CD19CAR and other surface markers can be kept expressed.

Provided herein are methods and compositions for treating a subject suffering from a deficiency in α-L-Iduronidase in the CNS. The methods include systemic administration of a bifunctional fusion antibody comprising an antibody to a human insulin receptor and an α-L-Iduronidase. A therapeutically effective systemic dose is based on the specific CNS uptake characteristics of human insulin receptor antibody-α-L-Iduronidase fusion antibodies as described herein.

The invention discloses expression of human insulin and analogues thereof in a methanol yeast containing dual promoters and a high-efficient preparation method of the human insulin and the analogues thereof, and the method is characterized in that recombinant human insulin and the analogues thereof are prepared by constructing an expression vector containing the dual promoters and engineering cells in host cells of the methanol yeast, and performing the optimal fermentation technology and the high-efficient purification technology, so that the production cost of the recombinant human insulin and the analogues thereof can be effectively reduced.

The invention discloses a culture medium used for culturing adipose mesenchymal stem cells, and applications thereof. The culture medium comprises insulin human, human serum albumin, transferring, fibronectin, ascorbic acid, biotin, PDGF, bFGF, and TGF-beta. It is confirmed by experiments that culturing with the culture medium is capable of obtaining a large amount of high quality mesenchymal stem cells, and the mesenchymal stem cells possess excellent stem cell characteristics, higher immunosuppression activity, and low immunogenicity, and are more suitable to be used for allogeneic cell therapy. The potential risk of clinical applications of exogenous animal serum is avoided, cell pollution rate is reduced, proliferation of human adipose mesenchymal stem cells is promoted, aging speed of human adipose mesenchymal stem cells in in-vitro culture is reduced. Requirements on large scale industrialized production of adipose mesenchymal stem cells needed by clinical therapy are satisfied, and problems such as unstable properties of different batches and high cost are solved at the same time.

The present invention utilizes transgenic tomato as bioreactor to produce human insulin. The human insulin is produced through designing plant preference codon, artificially synthesizing several DNAsegment, splicing, and adding endoplasmic reticulum residence sequence of KDEL in the C terminal to avoid the degradation of insulin in plant cell. The gene is transformed into tomato via agrobacterium mediating process under the driving of CaMV35S promoter and fruit specificity promoter 2A12 to express human insulin in tomato fruit. This kind of tomato capable of producing human insulin may become one kind of convenient oral product for preventing and treating some diabetes of depending insulin and autoimmune dysfunction, and may be used to extract human insulin for making injection.

The invention relates to the field of genetic engineering and discloses a pair of polypeptides and encoding genes thereof, with sequences shown as SEQ ID NO. 1 to 4. The pair of polypeptides is used for establishing a pair of a recombinant transcription activator like effector (TALE) and a transcription activator like effector nuclease (TALEN); the TALEN is a fusion protein formed by fusing a pair of DNA recognizing proteins with a DNA severing protein respectively; and the TALEN can perform targeted severing on a target site of a human insulin gene so as to achieve targeted modification on the human insulin gene, with the characteristics of strong specificity, high efficiency and high accuracy.

The invention discloses a serum-free medium for a monkey embryonic stem cell. The serum-free medium comprises a basic medium and additives, wherein the additives are dissolved in the basic medium; the basic medium comprises a DMEM-F12 medium; and the additive comprises L-glutamine, non-essential amino acid, L-ascorbic acid, sodium selenite, beta-mercaptoethanol, human serum albumin, heparin sodium, human transferring, insulin human, a basic fibroblast growth factor, a transforming growth factor beta 1 and an epidermal growth factor. The serum-free medium is capable of maintaining an undifferentiation state and totipotency of the monkey embryonic stem cell within a relatively long time, and is applicable to culture of the monkey embryonic stem cell in both a free-feeding layer system and a feeding layer system.

The invention discloses a serum substitute for cell culture. Each liter of the serum substitute comprises the following components: 140-165 mg of amino acid, 15-20 mg of asparagine, 15-25 [mu] M of VC, 70-110 [mu] M of vitamin H, 150-200 [mu] M of tocopheryl acetate, 80-120 [mu] M of tocopherol, 10-15 [mu] M of vitamin A, 15-25 g of a bovine pituitary extract, 80-120 [mu] g of an insulin-like growth factor I, 250-350 [mu] g of catalase, 450-550 [mu] M of insulin human recombinant, 5000-10000 [mu] M of human transferrin, 5000000U of superoxide dismutase, 5mM-20mM of corticosterone, 150000-250000mM of D-galactose, 100-150mM of diethanolamine hydrochloride, 49-165mM of glutathione, 80-120mM of l-carnitinechloride, 140.901-161.403mg of inorganic salts and 5-15g of Pluronic F-68.

The invention discloses a human umbilical cord mesenchymal stem cell serum-free culture medium and a preparation method and application thereof. The human umbilical cord mesenchymal stem cell serum-free culture medium comprises a basic culture medium, amino acid substances, trace elements, recombinant human serum albumin, transferrin, recombinant human insulin, recombinant human epidermal growth factors, alkaline fibroblast growth factors and buffer salt, wherein the amino acid substances comprise glycine, alanine, asparagine, aspartic acid, glutamic acid, proline, serine and glutamine. The serum-free culture medium is clear in components, stable in batch, free of serum, antibiotics and other serum analogues, and free of heterogeneous pollution and immunological rejection risks and safe touse; by the serum-free culture medium, normal cell adherence and good cell growth state can be kept; the serum-free culture medium has stemness and is suitable for the growth and stemness maintenanceof human umbilical cord mesenchymal stem cells.

A liquid supplying device for a human insulin injection includes a substrate, a liquid storage chamber, a flow-guiding-and-actuating unit, a sensor and a driving chip. The flow-guiding-and-actuating unit includes a liquid guiding channel having a liquid guiding outlet in fluid communication with a liquid storage outlet of the liquid storage chamber. The sensor contacts with the human skin to measure a blood glucose level contained in sweat. The driving chip is configured to control the actuation of the flow-guiding-and-actuating unit, control open / closed states of the switching valves and receive the measured data from the sensor for determination. By driving the flow-guiding-and-actuating unit, a pressure gradient is generated, and an insulin liquid stored in the liquid storage chamber is transported to the liquid guiding outlet through the liquid guiding channel, flowing into a microneedle patch, and injected into a subcutaneous tissue through a plurality of hollow microneedles.

The present invention is related to human insulin containing additional disulfide bonds and methods of making such.

The invention relates to a culture medium for adipogenic differentiation of mesenchymal stem cells and an application thereof. The culture medium is characterized by comprising a basic culture mediumand an additive component, wherein the culture medium comprises: a variety of components like inorganic salt, amino acid and vitamins which are optimally designed, and components like recombinant human serum albumin, sodium carboxymethylcellulose, 3-isobutyl-1-methylxanthine, recombinant human insulin, dexamethasone, pioglitazone hydrochloride, palmitic acid, oleic acid, linoleic acid and cholesterol. According to the invention, through addition and proportioning of optimally-designed components, the capacity of the culture medium for inducing mesenchymal stem cells to differentiate into adipose cells is obviously improved, and the differentiation time is further shortened.

The invention relates to the field of plant gene engineering and provides a sequence of a human insulin gene designed and synthesized according to plant preferred codons. A plant expression vector isbuilt for the synthetic human insulin gene, so that the gene can be transferred into tomatoes by an agrobacterium rhizogenes-mediated method under the drive of a CaMV 35S promoter and a fruit-specificpromoter 2A12 to express human insulin in tomato fruit. The tomatoes capable of making human insulin can be used as a convenient oral product for preventing and treating insulin-dependent diabetes mellitus and autoimmune disorder diabetes mellitus, and human insulin can be extracted from the tomatoes to be made into injection preparations.

The invention discloses a serum-free culture medium for mesenchymal stem cells. The serum-free culture medium for the mesenchymal stem cells comprises the following ingredients: fibronectin at the final concentration of 25mu g / ml, basic fibroblast growth factors at the final concentration of 10ng / ml, human epidermal growth factors at the final concentration of 15ng / ml, recombinant human insulin at the final concentration of 1mg / ml, human transferrin at the final concentration of 0.55mg / ml, human blood albumin in a volume ratio of 5 percent, sodium selenite at the final concentration of 0.67mug / ml, L-carnitine at the final concentration of 5mM and resveratrol at the final concentration of 30mu M. When the mesenchymal stem cells are cultured by the serum-free culture medium for the mesenchymal stem cells, animal-derived serum is not contained, so infection risks can be controlled; the L-carnitine and the resveratrol which are added into the serum-free culture medium for the mesenchymalstem cells can effectively improve the growth state of the mesenchymal stem cells; and the growth speed of the mesenchymal stem cells is remarkably improved, and the biological characteristics of themesenchymal stem cells are kept unchanged.

Compositions and methods are provided for screening for compounds that modulate insulin promoter activity. Vectors that express green fluorescent protein under the control of the human insulin promoter are introduced into mouse and human cells in which the insulin promoter is expressed in a glucose-responsive manner. Such cells are then used to screen for compounds that modulate insulin promoter activity.

The invention relates to a non-human transgenic mammal that is useful for the production of a protein of interest that may be toxic to the mammal. The mammal is characterized by the fact that it is transgenic for the production in its milk of an inactive form of the protein of interest, preferably recombinant human insulin. It is not possible to produce recombinant human insulin in transgenic mammals since this molecule has a certain degree of biological activity in the mammals and could be toxic to the mammal. Thus, the invention involves cloning a genetic construct comprising a sequence encoding a modified human insulin precursor under the control of a beta casein promoter in an expression vector. It also involves transfecting the expression plasmid into fetal bovine somatic cells, such as fibroblasts, and enucleating bovine oocytes by nuclear transfer to generate transgenic embryos. The invention gives rise to transgenic bovine that will be able to produce a modified human insulin precursor in their mammary glands. Afterwards, the milk of these transgenic mammals can be collected, the modified human insulin precursor can be converted in vitro into recombinant human insulin, and the recombinant human insulin can be purified to homogeneity as a pure biopharmaceutical product.

The invention was involved in expression of human insulin gene, especially for the expression of human insulin gene in Pichia Pastoris GS115 / pPICM#101 whose storage number wais CCTCC NO.M204071. It contained B and A strand synthesis, construction of express plasmid and engineering strain, transform of host cell, filter of transformant and over expression engineering strain, etc. The method left out the difficult procedure, reduced the process steps and shortened time, so the technique was simple. It overexpressed 10-100 folds than the others. It provided one new simple method for human insulin commercial process.

The invention relates to the biological field, in particular to a novel insulin derivative which is characterized in that the general formula of the compound is CnH2n + 1NH-X-D, wherein, n is equal to 14 to 22, CnH2n + 1NH is straight chain saturated fatty amino group; X is a natural amino acid except genetic coding proline or a 1-amino-1-carboxyl unnatural amino acid; D is a natural pig insulin stripped of B30 amino acid residue or a human insulin stripped of B30 amino acid residue. The insulin derivative of the invention is a novel derivative formed by connecting a fatty chain to the natural insulin or to the Alpha-carboxyl of the C-terminal residue on the parent B chain of the analogs. The preparation method is to take the natural pig insulin or human insulin as the starting material, cut off the first amino acid residue on the C terminal of the B chain of the insulin, and then take trypsin as the catalyst to cause the amino acid derivative connected with a fatty chain to become the novel insulin derivative connected with a fatty chain on the C terminal of B chain through enzymic reaction. The derivative has potential mouth taking and long-acting functions.

The invention relates to a serum-free medium of peripheral blood cells. The serum-free medium comprises a base medium and an additive component; the base medium is RPMI 1640 medium; the additive component is composed of, by volume, 35 to 50mg / L of interleukin-2, 1 to 2g / L of heparin, 25 to 30mg / L of transferring, 20 to 30mg / L of insulin human recombinant, 10 to 20mg / L of M-type phytohaemagglutinin, 7 to 10mg / L of L-glutamine, and 2 to 4g / L of sodium bicarbonate. Compared with the prior art, the serum-free medium of peripheral blood cells possesses following advantages: the serum-free medium of peripheral blood cells is capable of satisfying requirements of leukocyte rapid and large amount propagation in peripheral blood cells, is especially suitable to be used for culturing of peripheral blood cells, and possesses excellent propagation effect and cell survival rate.