37 results about "Human follicle stimulating hormone" patented technology

The invention provides recombinant native and mutein forms of human follicle stimulating hormone beta subunit (FSH beta) with characteristic glycosylation patterns which are influential in the metabolic activity of the protein. The invention also provides recombinant mutant forms of the human alpha subunit common to FSH, LH, CG, and TSH, to obtain hormones which also have unique glycosylation patterns. Also provided are recombinant materials to produce these subunits separately or together to obtain complete heterodimeric hormones of regulated glycosylation pattern.

The invention discloses a long-acting recombinant human follicle-stimulating hormone-Fc fusion protein (hFSH-Fc) and a preparation method thereof. The recombinant hFSH-Fc is a dimerization fusion protein. An amino acid sequence of the hFSH-Fc comprises an hFSHbeta subunit, CTP (carboxy-terminal peptide), an hFSHalpha subunit, a flexible peptide linker and a human IgG (immunoglobulin G)2 Fc variant from the N terminal to the C terminal in sequence. The hFSH-Fc has longer half-life in vivo and smaller side effects than existing hFSH. The invention also relates to an application of a recombinant hFSH-Fc composition to preparation of drugs for treating and/or preventing infertility.

The invention discloses a long-acting recombinant human follicle-stimulating hormone-Fc fusion protein (hFSH-Fc) and a preparation method thereof. The hFSH-Fc is a dimerization fusion protein. An amino acid sequence of the hFSH-Fc comprises an hFSHbeta subunit, CTP (carboxy-terminal peptide), an hFSHalpha subunit, a flexible peptide linker and a human IgG (immunoglobulin G) Fc variant from the N terminal to the C terminal in sequence. The hFSH-Fc has longer half-life in vivo and smaller side effects than existing hFSH. The invention also relates to an application of a recombinant hFSH-Fc composition to preparation of drugs for treating and/or preventing infertility.

The invention discloses a culture method for high-efficiency human follicle stimulating hormone expression CHO cells and belongs to the technical field of bio-pharmaceuticals. According to the culture method, seed cells are prepared into cell suspension, and subculture is performed after resuspension through a serum-free basal culture medium to obtain seed suspension; the obtained seed suspension is inoculated into a bioreactor to be cultured, the serum-free basal culture medium is fed at regular intervals in the culture process, growth situations of the cells are monitored, glucose solution is supplemented, and the cells are harvested after culture. According to the method, the serum-free basal culture medium is used throughout the process, culture in a fermentation tank is performed after reviving and amplification through bottle shaking, the process flow is simple, the cell culture density is high and can be as high as 4*107/mL , the cell survival rate reaches more than 98%, maintaining time under high density and a high survival rate is long, the yield of end products can reach 40mg/L, and the method is applicable to industrial production.

The invention discloses a long-acting recombinant human follicle-stimulating hormone-Fc fusion protein (hFSH-Fc) and a preparation method thereof. The hFSH-Fc is a dimerization fusion protein. An amino acid sequence of the hFSH-Fc comprises an hFSHbeta subunit, CTP (carboxy-terminal peptide), an hFSHalpha subunit, a flexible peptide linker and a human IgG (immunoglobulin G)2 Fc variant from the N terminal to the C terminal in sequence. The hFSH-Fc has longer half-life in vivo and better efficacy than existing FSH. The invention also relates to an application of a recombinant hFSH-Fc composition to preparation of drugs in the field of animal breeding.

The invention discloses a method and culture medium for producing a human follicle stimulating hormone through high-density perfusion culture of recombinant CHO cells and an application of the human follicle stimulating hormone, and belongs to the field of biological pharmacy. The preparation method comprises the steps of after CHO cell recovery, performing cell density amplification with a basicculture medium; after the density of living cells achieves a certain density, performing perfusion culture with a production culture medium; additionally adding a fresh production culture medium to the inner part of a bioreaction tank, trapping the CHO cells with a hollow fiber trapping device connected to the outer part of the reaction tank, so that the cells can maintain high density and vitality; and when the cells in the reactor are in the stable state, starting expression product collection of the CHO cells. Through bioreactor spinner culture and a circumscription cell trapping device, the cells can be trapped, broken slices and expressive protein can be filtered, and high-density environment can be provided for cell growth. Besides, through backward flushing of a hollow fiber column,the fiber plugging is reduced, the high-density culture time of the cells can be prolonged, and the batch yield can be notably increased.

The invention relates to anti-human follicle-stimulating hormone receptor antibodies. The invention also relates to use of the anti-human follicle-stimulating hormone receptor antibodies in the treatment of cancer. The present invention also relates to a process for detecting cancerous cells and a process for diagnosis cancer. The present invention finds an application in the therapeutic and diagnostic medical technical fields.

The present invention relates to recombinant human follicle-stimulating hormone and a preparation method and a drug application thereof. The recombinant human follicle-stimulating hormone is rhFSH heterodimer containing alpha and beta chains; for a mammalian cell CHO expression system, optimized design of coding genes is performed; and expression plasmids are constructed in a targeted manner, so that expression and correct combination of the alpha and beta chains of the hFSH in mammalian cells CHO are better facilitated. The rhFSH has the advantages of simple process, high target product yield, high purity, high in vivo activity, convenience for intermediate process control, easiness for large-scale production and the like.

The invention provides recombinant hFSH-Fc fusion protein. The fusion protein is dimerized fusion protein, wherein one monomer amino acid sequence sequentially comprises an hFSH beta sub-unit, CTP, CTP and an hFSH alpha sub-unit from the end N to the end C, and the other monomer amino acid sequence sequentially comprises CTP, an hFSH alpha sub-unit, an hFSH beta sub-unit and CTP from the end N tothe end C. The two pieces of CTP are connected to an encoding sequence of an FSH, regions important to receptor binding or bio-activity are avoided, the affinity of receptor binding is not affected, immunogenicity is avoided, and the volume lifetime can be remarkably prolonged. Meanwhile, the in-vivo half-life period of FSH-CTP is longer, the ending half-life period is 2-3 times that of recombinant FSH-WT, one time of injection per week can replace seven times of injection every day, and the bio-activity is higher.

A nucleic acid sequence is provided that encodes a chimeric protein comprising a ligand that comprises a naturally occurring or modified follicle stimulating hormone sequence, e.g., an FSHp sequence, or fragment thereof, which ligand binds to human follicle stimulating hormone (FSH) receptor, linked to either (a) a nucleic acid sequence that encodes an extracellular hinge domain, a transmembrane domain, a co-stimulatory signaling region, and a signaling endodomain; or (b) a nucleic acid sequence that encodes a ligand that binds to NKG2D. The vector containing the nucleic acid sequence, the chimeric proteins so encoded, and modified T cells expressing the chimeric protein, as well as method of using these compositions for the treatment of FSHR-expressing cancers or tumor cells are also provided.

The invention discloses an analytical method of utilizing adherent cell-human ovarian granular cell tumor cell strain (KGN) to evaluate in-vitro biological activity of human follicle stimulating hormone. The method comprises the following steps: using the human follicle stimulating hormone to stimulate the human ovarian granular cell tumor cell, measuring the content of generated progesterone, so as to reflect the activity of the human follicle stimulating hormone. The analytical method of using the human ovarian granular cell tumor adherent cell to evaluate the in-vitro biological activity of the human follicle stimulating hormone, disclosed by the invention, is effective, stable and quick, and avoids the limitations of long time consumption, complicated operation and poor result repeatability in the animal in-vivo ovarian weight increment method; meanwhile, the progesterone content is determined by a commercial enzyme linked immunosorbent assay (ELISA) kit, so that the biological activity of the human follicle stimulating hormone can be reflected truly and more accurately.

The present invention relates to a long-acting human follicle-stimulating hormone formulation having improved in vivo duration and stability, comprising a human follicle-stimulating hormone conjugate that is prepared by covalently linking human follicle-stimulating hormone with an immunoglobulin Fc region via a non-peptidyl polymer, and a preparation method thereof. The long-acting human follicle-stimulating hormone formulation of the present invention maintains in vivo activity of human follicle-stimulating hormone at a relatively high level and remarkably increases the serum half-life thereof.

Provided is an aqueous formulation of a human follicle stimulating hormone (hFSH) which is stabilized to maintain the activity of hFSH for a prolonged period of time. The formulation of the present invention is an aqueous formulation comprising a therapeutically effective amount of hFSH stabilized in a phosphate buffer containing glycine, methionine and a non-ionic surfactant, preferably polysorbate 20, which is capable of maintaining the activity of hFSH for an extended period of time.

The invention discloses a recombinant human follicle stimulating hormone and a preparation method thereof. The preparation method comprises the following steps: jointly transfecting a constructed expression vector containing an FSHalpha subunit and an expression vector containing an FSHbeta subunit into Chinese hamster ovary cells, screening out stably expressed cell strains, and purifying and culturing the cell strains to obtain a culture solution, thereby obtaining the recombinant human follicle stimulating hormone, wherein the C-terminus of the FSHbeta subunit is connected with CTP. According to the preparation method of the recombinant human follicle stimulating hormone, hFSHalpha and hFSHbeta sequences comprising introns are cloned from human embryonic kidney 293 cell genomes respectively, CTP sequences are connected to the terminus of the beta subunit in a primer extension mode to obtain hFSHbeta-CTP, then the two subunit sequences are transfected to CHO-S cells, rhFSH-CTP is obtained through expression, the half-life period of the rhFSH-CTP obtained by expression in vivo is longer, the final half-life period is 2-3 times that of recombinant FSH, and the biological activity is higher.

The invention provides a double-antibody sandwich enzyme-linked immunoassay method for quantitatively detecting human follicle-stimulating hormone (hFSH). The invention mainly aims to solve the technical problems of low sensitivity, low system stability, limited detection scope, low repeatability, complex operation, and the like, in the prior art. According to the invention, two paired specific monoclonal antibodies (namely, anti-FSH alpha antibody and anti-FSH beta antibody) are adopted for detecting and quantizing human follicle-stimulating hormone or related follicle-stimulating hormone sample. The method provided by the invention has the advantages of easy operation, high sensitivity, high specificity, high system stability, wide FSH detection scope and capability of directly reflecting the concentration or in vitro biologic activity of an FSH standard substance or sample according to a light absorption value (OD value) detected by a microplate reader.