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WO1993020199A1 - Recepteur de gonadotrophine humaine (recepteur hfs) - Google Patents

Recepteur de gonadotrophine humaine (recepteur hfs) Download PDF

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Publication number
WO1993020199A1
WO1993020199A1 PCT/EP1993/000780 EP9300780W WO9320199A1 WO 1993020199 A1 WO1993020199 A1 WO 1993020199A1 EP 9300780 W EP9300780 W EP 9300780W WO 9320199 A1 WO9320199 A1 WO 9320199A1
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Prior art keywords
leu
ser
fsh
hfsh
ala
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PCT/EP1993/000780
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English (en)
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WO1993020199A9 (fr
Inventor
Rein Dijkema
Renato De Leeuw
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Akzo Nobel N.V.
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Publication of WO1993020199A1 publication Critical patent/WO1993020199A1/fr
Publication of WO1993020199A9 publication Critical patent/WO1993020199A9/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
    • C07K14/723—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2869—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against hormone receptors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence

Definitions

  • the invention relates to the field of reproductive medicine, particularly human reproductive medicine.
  • Gonadotropins follicle stimulating hormone (FSH) , chorionic gonadotropin (CG) , luteinizing hormone (LH) and thyroid stimulating hormone (TSH)
  • FSH follicle stimulating hormone
  • CG chorionic gonadotropin
  • LH luteinizing hormone
  • TSH thyroid stimulating hormone
  • the present invention provides such a cell. It also provides a novel gonadotropin receptor, i.e. the human follicle stimulating hormone receptor.
  • the DNA sequence is useful for studying the characteristics of the receptor by site directed mutations, thereby enabling to elucidate the parts of the receptor involved in the various aspects of its functions.
  • the amino acid sequence can be used to produce synthetic peptides in order to identify the smallest peptide still having binding affinity for FSH.
  • polypeptides comprising the extracellular part of the FSH receptor can be constructed.
  • the peptides and polypeptides of the invention can also be used directly as competing compounds for the endogenous receptors or in diagnostic test kits testing for the ligand for the receptor.
  • Antibodies or antiserum directed against a polypeptide according to the invention have use in diagnostic immunoassay's and generation of anti- idiotype antibodies.
  • a more preferred use is the use as an antidote against oversti ulation with FSH. This occurs regularly in IVF (in vitro fertilization) protocols resulting in ovarian hyperstimula ion.
  • the antibodies may be formulated into pharmaceutical formulations by mixing with suitable pharmaceutical acceptable carriers in a manner known to those skilled in the art.
  • a specific polypeptide according to the invention in any of the embodiments described above can be used to produce antibodies, both polyclonal, monospecific and monoclonal. Such use of a polypeptide according to the invention and such an antibody fall within the scope of the invention.
  • polyclonal antibodies When polyclonal antibodies are desired, techniques for producing and processing polyclonal sera are known in the art (e.g. Mayer and Walter, eds, Immunochemical Methods in Cell and Molecular Biology, Academic Press, London, 1987) .
  • a selected mammal e.g. a rabbit is given (multiple) injections with one of the above-mentioned immunogens, e.g. corresponding to about 20 ⁇ g to about 80 ⁇ g of polypeptide per immunization. Immunization is carried out with an acceptable adjuvant, generally in equal volumes of immunogen and adjuvant.
  • Acceptable adjuvants include Freund's complete, Freund's incomplete, alum-precipitate or water-in-oil emulsions, with a preference for Freund's complete adjuvant for the initial immunization. For booster immunization Freund's incomplete adjuvant is preferred.
  • the initial immunization consists of the administration of approximately 1 ml emulsion at multiple subcutaneous sites on the backs of the rabbits. Booster immunizations utilizing an equal volume of immunogen are given at about one monthly intervals and are continued until adequate levels of antibodies are present in an individual rabbits serum. Blood is collected and serum isolated by methods known in the art.
  • Monospecific antibodies to each of the immunogens are affinity purified from polyspecific antisera by a modification of the method of Hall et al. (Nature 311, 379-387 1984) , prepared by immunizing rabbits as described above with the purified proteins.
  • Monospecific antibody as used herein is defined as a single antibody species or multiple antibody species with homogeneous binding characteristics for the relevant antigen.
  • Homogeneous binding as used herein refers to the ability of the antibody species to bind to a specific antigen or epitope.
  • Monoclonal antibody reactive against one of the above- mentioned immunogens can be prepared by immunizing inbred mice, preferably Balb/c with the appropriate protein by techniques known in the art (Kohler and Milstein, Nature 256; 495-497, 1975). Hybridoma cells are subsequently selected by growth in hypoxanthine, thy idine and a inopterin in an appropriate cell culture medium such as Dulbecco's modified Eagle's medium (DMEM) . Antibody producing hybridomas are cloned, preferably using the soft agar technique of MacPherson, (Soft Agar Techniques, Tissue Culture Methods and Applications, Kruse and Paterson, eds. , Academic Press, 276, 1973) .
  • Discrete colonies are transferred into individual wells of culture plates for cultivation in an appropriate culture medium.
  • Antibody producing cells are identified by screening with the appropriate immunogen.
  • Immunogen positive hybridoma cells are maintained by techniques known in the art.
  • Specific anti-monoclonal antibodies are produced by cultivating the hybridomas in vitro or preparing ascites fluid in mice following hybridoma injection by procedures known in the art.
  • the polyclonal or monoclonal antibodies can show agonistic or antagonistic activity as compared to the activity of the natural ligand.
  • Specific antagonistic or agonistic activity may involve the interference of the antibodies with the signal transduction system in the cells expressing the FSH receptor, without competing for the binding of FSH to its receptor.
  • the DNA coding for the receptor or for an altered receptor can be inserted into a suitable vector for expression in either a prokaryotic or a eukaryotic host.
  • the hosts may be bacteria, phages, yeasts, funghi, animal cells or plant cells, preferred however are mammalian cells.
  • the vector into which the DNA is inserted may be any suitable vector. It may comprise suitable regulating elements such as promoters, enhancers, repressors, etc. If necessary, it may also comprise a signal sequence to transport the protein translated from the DNA to the surface or the outside of the host, even though the receptor does contain its own signal sequence.
  • a cell which preferably expresses no related receptors and more preferably no other receptors at all, and which cell expresses the receptor according to the invention in altered or unaltered form, in its (outer) membrane.
  • a signal producing system in the cell is needed.
  • the signal producing system may be the cell's own or may be cotransfected with the DNA coding for the receptor.
  • the signal will be provided by the so called second messenger system, which works through G- proteins which are associated with the receptor at its surface site. Any other signal producing system as well as second messenger initiated test system will be suitable, as long as the signal is somehow measurable.
  • Oligodeoxynucleotides were synthesized using the phosphoramidite method on an Applied Biosystems 381A DNA synthesizer. They correspond to nucleotide position 1237-1255 (transmembrane region II) and 1843-1861 (transmembrane region VII) of the hFSH-R (Minegish et al., Bioch.Bioph.Res.Comm. 175, 1125, 1991) . Both oligonucleotides were used as primers in a polymerase chain reaction (Maniatis et al. , Cold Spring Harbor Laboratory, "Molecular Cloning: A Laboratory Manual", 1989) to generate a 625bp DNA fragment amplified from human genomic DNA (Clonetech) . The resulting product from PCR was verified by DNA sequence analysis (Pharmacia,T7-sequencing kit) .
  • the partial hFSH-R amplification product obtained by PCR (section 1.1.) was used as probe (Pharmacia, oligolabelling kit) to screen 3.6xl0 5 recombinant phages of the human testis cDNA library (section 1.2.) • Prehybridization of filters was carried out for 5 hours at 65 °C in a solution containing 6xSSC (0.9 mol/1 NaCl,0.09 mol/1 Na-citrate, pH 7.0), lOxDenhardt (lxDenhardt:200 ⁇ g/ml Ficoll-70 (Pharmacia), 200 ⁇ g/ml polyvinylpyrrolidone (Sigma) , 200 ⁇ g/ml bovine serum albumin (BSA, Sigma) , 50 ⁇ g/ml sheared and denatured herring sperm DNA (Sigma) , 9% dextran sulphate (Pharmacia), and 0.1% sodium dodecyl sulphate (
  • Hybridization was performed in the same solution by the addition of the 32 P-labelled DNA probe (2.3xl0 5 cpm/ml) . The mixture was incubated overnight at the same temperature. Filters were washed in solutions with decreasing salt concentrations (O.lxSSC, 65 °C) . Positive recombinant phages were purified by two successive rounds of phage titration and hybridization. Phage DNA inserts were isolated and subcloned in the endoR EcoRI site of pGEM3Z (Promega) and characterized by endoR mapping and DNA sequence analysis.
  • CHO cells (CHO Kl) were obtained from ATCC (CCL61) . They were cultured in M505 medium that consisted of a mixture (1:1) of Dulbecco's Modified Eagle's Medium (DMEM, Gibco 074-2100) and Nutrient mixture F12 (Ham's F12, Gibco 074-1700) supplemented with 2.5 mg/ml sodium bicarbonate (Baker), 55 ⁇ g/ml sodium pyruvate (Fluka) , 2.3 ⁇ g/ml 3-mercaptoethanol (Baker), 1.2 ⁇ g/ml ethanolamine (Baker), 360 ⁇ g/ml L- glutamine (Merck) , 0.45 ⁇ g/ml sodiumselenite (Fluka) , 62.5 ⁇ g/ml penicillin (Mycopharm) , 62.5 ⁇ g/ml streptomycin (Serva) , and 10% fetal calf serum (FCS, Bocknek) .
  • Recombinant constructs used for transformation of CHO cells consist of the expression vector pKCRhFSH-R (section 2.1.) and the selection vector pAG60MT2.
  • the latter vector was constructed by insertion of a 3kb human MTII ⁇ -containing endoR Hindlll fragment (Karin and Richards, Nature 299, 797, 1982) into the endoR Hindlll site of pAG60 (Colbere-Garapin et al., J.Mol.Biol 150, 1, 1981) ; the transcription of this gene was directed towards the tk-promoter that was located in front of the neomycin resistence gene.
  • CHOhFSH- RlCdlO refers to transformed CHO cells obtained after successive selection by neomycin and 10 ⁇ mol/1 CdCl 2 .
  • CHOhFSH-RlCdlO cells pellet were homogenised with a teflon glass homogeniser in ice-cold 10 mmol/1 Tris- HCl buffer, pH 7.4, supplemented with 0.25 mol/1 sucrose and 5 mmol/1 MgCl2- The homogenate was diluted to 2.5 x IO 6 cells (starting material)/ml with homogenisation buffer. For saturation experiments, aliquots homogenate (200 ⁇ l/tube) were incubated with increasing concentrations 125 I-hFSH (200 ⁇ l/tube; 4-150 pmol/1) with or without excess unlabeled recFSH (10 IU/100 ⁇ l/tube) in polypropylene microfuge tubes.
  • the assay buffer consisted of 10 mmol/1 Tris-HCl, pH 7.4, supplemented with 5 mmol/1 MgCl2 and 1 g/1 bovine serum albu ine (BSA; Sigma, St Louis, MO, USA) . After 24 h of incubation at room temperature (RT) , 500 ⁇ l ice-cold assay buffer was added, and bound and free hormone were separated by centrifugation (5 in at 15.000 x g) . The bound radioactivity was measured with a LKB gamma counter. The ⁇ (equilibrium dissociation constant) and &___ (maximum binding capacity) were assessed by Scatchard plot analysis. Competition of antisera for binding was assessed using the single cell clone mentioned previously expressing the human FSH-R, selected by subsequent exposure to G418 (800 ⁇ g/ml; Geneticin;
  • control serum that is included is a serum of saline-injected mice. The results are presented in
  • the CHOhFSH-RlCdlO cells were washed once, resuspended in M505, pH 7.4, supplemented with 10% FCS, and cultured in 24 well-plates (Nunclon; 0.1% gelatin coated) at a concentration of 2 x IO 5 cells/ml/well for 48 h at 37 "C in a humidified atmosphere of 5% C0 /95% 0 2 .
  • the remaining cells were treated with 0.5 ml 1-propanol which was followed by ultrasonification for 2 min. Subsequently, the content of each well was transfered to Eppendorf tubes and stored at -20°C.
  • a cAMP (RIANEN) kit NNN
  • the cellular samples were lyophilized using a Speed Vac and reconstituted in 0.5 ml kit buffer. The medium samples were measured directly using a callibration curve of standard cAMP in CHO cell culture medium.
  • GST-FSH-R 1 contains a 1000 basepair fragment coding for the complete extracellular N-terminus of the FSH- R.
  • Fusion proteins 2 and 3 contain only parts of this sequence: fusion protein 2 (GST-FSH-R 2) contains a 600 basepair fragment coding for the N-terminal part of the FSH-R present in GST-FSH-R 1; fusion protein 3 (GST-FSH-R 3) contains a 400 basepair fragment coding for the C-terminal part of the FSH-R present in GST- FSH-R 1) . These fragments were derived by polymerase chain reaction (PCR) on the FSH-R cDNA clones ( Figure 2A; seq. ID No. 1) . Via this procedure restriction sites were introduced, which facilitated subsequent cloning procedures.
  • PCR polymerase chain reaction
  • GST-FSH-R 1 we used the primers 5 » -TGTCATCATCGGATC-3' and 5'-TCTGAGGATGTTGTAC- 3• ; for GST-FSH-R 2 we used the primers 5'- TGTCATCATCGGATC-3' and 5'-AGGCAGGGAATGGATCC-3 ' ; for GST-FSH-R 3 we used the primers 5'-AGAACAAGGATCC-3' and 5'-TCTGAGGATGTTGTAC-3' .
  • Some primers were complementary to the FSH-R cDNA sequence (SEQ ID N0:1). Only the complementary sequence of the primers is shown.
  • the FSH-R fragments were cloned into one of the pGEX expression vectors (Smith and Johnson, Gene 67, 31, 1988) . Depending on the reading frame, either pGEX-1 (for the 1000 bp and 600 bp FSH-R fragments) or pGEX-3X (for the 400 bp FSH-R fragment) were used. In order to study expression of the fusion proteins, these constructs were transformed into E. coli MC1061.
  • IPTG isopropyl-3-D- thiogalactopyranoside
  • Triton-X-100 was added to a final concentration of 1%, incubation was continued for 10 minutes on ice, and cells were sonified. The supernantant of the sonicate was incubated with gluthathione-agarose carrier (Pharmacia) for 30 minutes at 4°C. Carriers were washed several times with PBS, and finally with 50 mM Tris pH 8. The fusion proteins thus purified were used to generate antisera.
  • mice Six weeks old female BALB/c mice were injected intraperitoneally with 50 ⁇ g of bacterial fusion proteins in complete Freund's adjuvant. In addition,
  • mice were immunized with membranes of 10' CHO cells transfected with the human FSH-R.
  • an FSH-R membranes of 10' CHO cells transfected with the human FSH-R.
  • FSH-R single cell clone was used that was obtained by subsequent G418 (800 ⁇ g/ml; Geneticin, Gibco) and cadmium (2.5 ⁇ M) selection. Two subsequent intramuscular injections were given at three weeks intervals with 50 ⁇ g fusion protein or membranes of
  • mice were boosted intraperitoneally with 100 ⁇ g fusion protein or membranes of 10 7' cells in PBS. Four days after this final boost, sera and spleens were collected.
  • Erythrocyte depleted spleen cells were prepared according to Steenbakkers et al. (J. Imm. Methods 152,
  • Denatured and reduced protein preparations were applied on sodium dodecylsulfate (SDS) polyacrylamide gels and blotted onto nitrocellulose filters.
  • SDS sodium dodecylsulfate
  • small (approximately 200 ng) and equivalent amounts of protein were applied to the gels.
  • Membranes were blocked with 20% FCS, and incubated for 2 hours at room temperature with different concentrations of antibody preparations. Blots were washed with Tris-buffered saline (TBS) - Tween and incubated with Goat-anti-mouse alkaline phosphatase (AP) conjugate (Promega) for 45 minutes at room temperature.
  • TBS Tris-buffered saline
  • AP Goat-anti-mouse alkaline phosphatase conjugate
  • AP substrate nitrogen tetrazoline (NBT) / 5-bromo-4-chloro-3- indolyl phosphate (BCIP) ) reactions were done in 100 mM NaCl, 100 mM Tris pH 9.5, and 10 mM MgCl2) .
  • CHO cells were attached to the bottom of 24 well plates (approximately 10" cells/well) for three days. Cells were washed in phosphate buffered saline (PBS) for 10 minutes. After fixation with 4% paraformaldehyde in PBS for one hour at room temperature, the cells were washed in PBS containing 0.05% normal swine serum (NSS) and 0.02% Triton-X-100 for 10 minutes. After blocking with NSS in PBS-Triton for 1 hour, cells were incubated with antisera for 16 hours at room temperature.
  • PBS phosphate buffered saline
  • NSS normal swine serum
  • Triton-X-100 Triton-X-100
  • Antibodies raised against the human FSH-R were characterized by Western immunoblotting, immunostam . i.ng, and by thei.r competi.ti.on for binding and cAMP generation.
  • Antisera raised against the bacterial GST-FSH-R fusions recognized these proteins on Western blots (Fig. 6C) . Often, these antisera also recognized GST alone (control lanes) . After preclearing with GST, antisera specifically reacted with the FSH-R part of the fusion proteins. A representative example is shown in Figure 6B.
  • the anti GST-FSH-Rl serum inhibited cAMP generation to the same degree at the anti GST-FSH-R3 serum (Fig. 8) .
  • the GST-FSH-Rl antiserum competed less than the anti GST-FSH-R3 serum for 125 i- FSH binding (Fig. 7) .
  • These antibodies are potentially interesting as antagonists since they do not have to compete with the high affinity FSH binding in order to block the biological effect of FSH.
  • TCT AAC AGG GTT TTT CTC TGC CAA GAG AGC AAG GTG ACA GAG ATT CCT 207 Ser Asn Arg Val Phe Leu Cys Gin Glu Ser Lys Val Thr Glu lie Pro
  • CAA GCC CAA ATT TAT AGG ACA GAA ACT TCA TCC ACT GTC CAC AAC ACC 2079 Gin Ala Gin He Tyr Arg Thr Glu Thr Ser Ser Thr Val His Asn Thr 650 655 660 665
  • MOLECULE TYPE protein
  • Fig. 1 Physical maps of three hFSH-R cDNA clones and their position on their respective plasmids.
  • Fig. 2A Nucleotide sequence of the hFSH-R cDNA cloned in pGEM3Zcl (seq. ID No. 1) .
  • Fig. 2B Amino acid sequence of the hFSH-R coded for by the hFSH-R cDNA in pGEM3Zcl (seq. ID No. 2) .
  • Fig. 3 Binding of 125 I-hFSH to hFSH-R of a neomycin- CdC12 (10 ⁇ mol/1) selected CHO pool. Cell membranes were incubated with increasing concentrations 125 ⁇ - hFSH in the absence or presence of excess unlabeled recFSH. The saturation curve of specifically bound 125 I-hFSH together with the derived Scatchard plot are shown. The values represent the mean of duplicate determinations. The calculated Kd and B j ⁇ x are presented in the Scatchard plot (insert) .
  • Fig. 4 Dose dependent stimulation of intracellular cAMP by recFSH. The stimulations were repeated at different times of incubation. Values represent the mean of duplicate determinations.
  • Fig. 5 Dose dependent stimulation of extracellular (medium) cAMP by recFSH. The stimulations were repeated at different times of incubation. Values represent the mean of duplicate determinations.
  • FIG. 6A Western blot of GST-hFSH-R fusion proteins stained with Coomassie Blue.
  • Fig. 6B Western blot of GST-hFSH-R fusion proteins stained with antiserum raised against GST-hFSH-R3 fusion protein. The antiserum has been preclaered with GST proteins.
  • Fig. 6C Western blot of GST-hFSH-R fusion proteins stained with antiserum raised against GST-hFSH-Rl fusion protein.
  • Fig. 7 Interference of hFSH-R antisera with binding of 125j_ FSH to CH0 ce ⁇ s expressing the human F S H receptor (hFSH-R) .
  • Fig. 8 Interference of hFSH-R antisera with FSH-induced cAMP generation.

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Abstract

Nouveau récepteur de gonadotrophines, séquences d'acides aminés de ce récepteur, séquences d'acide nucléique codant pour ledit récepteur, hôtes recombinés comportant un tel acide nucléique, et dosages de dépistage utilisant ledit récepteur ainsi que des ligands tels que les anticorps dressés contre ce récepteur. Dans un mode préféré de réalisation, on a prévu des anticorps dressés contre le récepteur de l'hormone folliculostimulante humaine. Ces composés et procédés trouveront application dans le domaine de la médecine de la reproduction, notamment celle de l'homme.
PCT/EP1993/000780 1992-03-30 1993-03-29 Recepteur de gonadotrophine humaine (recepteur hfs) WO1993020199A1 (fr)

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EP92200886 1992-03-30
EP92200886.7 1992-03-30

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WO1993020199A1 true WO1993020199A1 (fr) 1993-10-14
WO1993020199A9 WO1993020199A9 (fr) 1994-01-06

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000073416A1 (fr) * 1999-05-28 2000-12-07 University Of Cincinnati Oligonucleotides antisens regulateurs de la fecondite et du cycle menstruel, et a usage chimiopreventif et chimiocuratif
WO2001088127A3 (fr) * 2000-05-18 2002-09-19 Bayer Ag Régulation du récepteur couplé à la protéine g analogue à l'hormone folliculo-stimulante humaine
EP0950711A3 (fr) * 1998-02-06 2003-09-17 Akzo Nobel N.V. Récepteurs de Gonadotropine
WO2018044929A1 (fr) * 2016-08-29 2018-03-08 David Weiner Compositions immunogènes consensus synthétiques optimisées ciblant le récepteur d'hormone de stimulation folliculaire (fshr)
WO2020033797A1 (fr) * 2018-08-09 2020-02-13 The Wistar Institute Anticorps anti-récepteurs de l'hormone de stimulation anti-inflammatoire
WO2022116853A1 (fr) * 2020-12-04 2022-06-09 康威(广州)生物科技有限公司 Préparation et utilisation d'un anticorps anti-fshr et d'un conjugué anticorps-médicament de celui-ci

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US4921808A (en) * 1986-06-25 1990-05-01 The Albany Medical College Of Union University Method for determining follicle stimulating hormone
WO1990013643A2 (fr) * 1989-05-05 1990-11-15 Genentech, Inc. Molecules receptrices d'hormone de glycoproteine
WO1992016620A1 (fr) * 1991-03-15 1992-10-01 Applied Research Systems Ars Holding N.V. Recepteur de l'hormone folliculostimulante humaine

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US4652450A (en) * 1984-08-31 1987-03-24 Research Corporation Bacterial FSH binding inhibitor
US4921808A (en) * 1986-06-25 1990-05-01 The Albany Medical College Of Union University Method for determining follicle stimulating hormone
WO1988008719A1 (fr) * 1987-05-12 1988-11-17 Aphton Corporation Immunocontraceptifs specifiques d'hormones de reproduction et leurs procedes d'utilisation
WO1990013643A2 (fr) * 1989-05-05 1990-11-15 Genentech, Inc. Molecules receptrices d'hormone de glycoproteine
WO1992016620A1 (fr) * 1991-03-15 1992-10-01 Applied Research Systems Ars Holding N.V. Recepteur de l'hormone folliculostimulante humaine

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Title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS. vol. 175, no. 3, 29 March 1991, DULUTH, MINNESOTA US pages 1125 - 1130 Minegish, Takashi; Nakamura, Kazuto; Takakura, Yumi; Ibuki, Yoshito; Igarashi, Masao 'Cloning and sequencing of human FSH receptor cDNA' *
ENDOCRINONOLGY vol. 126, no. 3, March 1990, pages 1318 - 1326 Dattatreyamurty B;Zhang SB;Reichert LE Jr; 'Polyclonal antibodies against follitropin (FSH) receptor interfere with hormone binding, but mimic the effects of FSH.' *
MOLECULAR ENDOCRINOLOGY vol. 4, 1990, pages 525 - 530 Sprengel, Rolf; Braun, Thomas; Nikolics, Karoly; Segaloff, Deborah L.; Seeburg, Peter H. 'The testicular receptor for follicle stimulating hormone: structure and functional expression of cloned cDNA' *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0950711A3 (fr) * 1998-02-06 2003-09-17 Akzo Nobel N.V. Récepteurs de Gonadotropine
WO2000073416A1 (fr) * 1999-05-28 2000-12-07 University Of Cincinnati Oligonucleotides antisens regulateurs de la fecondite et du cycle menstruel, et a usage chimiopreventif et chimiocuratif
US7273932B1 (en) 1999-05-28 2007-09-25 The University Of Cincinnati Antisense oligonucleotides for fertility and menstrual cycle regulation and for chemopreventive and chemotherapeutic use
WO2001088127A3 (fr) * 2000-05-18 2002-09-19 Bayer Ag Régulation du récepteur couplé à la protéine g analogue à l'hormone folliculo-stimulante humaine
WO2018044929A1 (fr) * 2016-08-29 2018-03-08 David Weiner Compositions immunogènes consensus synthétiques optimisées ciblant le récepteur d'hormone de stimulation folliculaire (fshr)
CN110121354A (zh) * 2016-08-29 2019-08-13 宾夕法尼亚大学理事会 靶向卵泡刺激素受体(fshr)的最优化的合成共有免疫原性组合物
US11865166B2 (en) 2016-08-29 2024-01-09 The Trustees Of The University Of Pennsylvania Nucleic acid encoding optimized immunogenic peptide that targets follicle stimulating hormone receptor
WO2020033797A1 (fr) * 2018-08-09 2020-02-13 The Wistar Institute Anticorps anti-récepteurs de l'hormone de stimulation anti-inflammatoire
US20210340263A1 (en) * 2018-08-09 2021-11-04 The Wistar Institute Anti-Follicule Stimulating Hormone Receptor Antibodies
WO2022116853A1 (fr) * 2020-12-04 2022-06-09 康威(广州)生物科技有限公司 Préparation et utilisation d'un anticorps anti-fshr et d'un conjugué anticorps-médicament de celui-ci

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