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WO1998048014A1 - Marqueur utilise dans le diagnostic de cancers - Google Patents

Marqueur utilise dans le diagnostic de cancers Download PDF

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Publication number
WO1998048014A1
WO1998048014A1 PCT/GB1998/001184 GB9801184W WO9848014A1 WO 1998048014 A1 WO1998048014 A1 WO 1998048014A1 GB 9801184 W GB9801184 W GB 9801184W WO 9848014 A1 WO9848014 A1 WO 9848014A1
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WO
WIPO (PCT)
Prior art keywords
protein
expressed
analogue
antibody
immunogenic fragment
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Application number
PCT/GB1998/001184
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English (en)
Inventor
Wilhelm Schwaeble
Original Assignee
University Of Leicester
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Publication date
Application filed by University Of Leicester filed Critical University Of Leicester
Priority to AU70677/98A priority Critical patent/AU7067798A/en
Publication of WO1998048014A1 publication Critical patent/WO1998048014A1/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
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4727—Mucins, e.g. human intestinal mucin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention relates to a cancer marker, agents specific against same and the therapeutic and diagnostic uses of same.
  • Mucins high molecular-weight glycoproteins
  • the typically high content of carbohydrates can account for up to 80% of the molecular weight of a mucin.
  • These carbohydrates are mainly O-linked oligosaccharides bound to serine and threonine residues in the protein backbone by a terminal N-acetylgalactosamine residue.
  • Abundant amino acids in mucins include threonine, serine, proline and frequently glycine and alanine.
  • seven different human mucins have been described (see for example Lesuffleur, T. etal, 1995, J. Biol. Chem.
  • mucin antigens Numerous alterations of mucin antigens have been described in neoplastic epithelial tissues and in the sera of patients with pancreatic breast, ovarian, and colon cancer (Ho, S.B et al, 1993, Cancer Res., 53.: 641-651). These antigens (DF3, CA 19-9, CA125, SPan-1, and DuPan 2) have been used as diagnostic tumour markers. In an experimental animal model, mucin production by human colon cancer cell lines positively correlates with the metastatic potential of a cell line. Alterations of mucin-type glycoproteins may contribute to changes in cellular adhesion and immune recognition.
  • the mucin MUC 1 has been established as a marker for diagnosis, prognosis and monitoring of breast cancer.
  • a problem with MUC1 is that it is also expressed by normal breast epithelial cells and this can give rise to false positive indications, or can raise doubts about actual positive indications.
  • carcinomas particularly carcinomas such as breast, ovarian and epidermal carcinomas in general, but not in normal cells.
  • the present invention provides a novel marker (termed MUC-B1) which overcomes the problems of the prior art.
  • a protein encoded by a cDNA having at least the sequence of SEQ ID NO: 1 (pBl) or a partially modified form thereof.
  • It may have the amino acid sequence of SEQ ID NO: 2 (ORF1) or a partially modified form thereof, or the sequence of SEQ ID NO: 3 (ORF2) or a partially modified form thereof.
  • partial modification and “partially modified” is meant, with reference to amino acid sequences, a partially modified form of the molecule which retains substantially the properties of the molecule from which it is derived, although it may of course have additional functionality. Partial modification may, for example, be by way of addition, deletion or substitution of amino acid residues. Substitutions may be conserved substitutions.
  • a partially modified molecule may be a homologue of the molecule from which it was derived. It may, for example, have at least 40% homology with the molecule from which it was derived. It may for example have at least 50, 60, 70, 80, 90 or 95% homology with the molecule from which it was derived.
  • An example of a homologue is an allelic mutant.
  • nucleotide sequences encoding the molecules or amino acid sequences may be partially modified to code for any such modifications to an amino acid sequence or molecule.
  • Nucleotide sequences may also of course be modified such that they still code for the same amino acid residues but have a different nucleotide sequence.
  • the protein is not expressed by normal cells such as breast and ovarian epithelium but is expressed by a substantial proportion of cancers, particularly epithelial cancers, such as breast, ovarian and colonic carcinomas. It may be expressed by at least for example 50, 60, 70, 80 or 90% of cancers.
  • a study showed it to be expressed in approximately 60% of specimens of breast carcinomas and 85% of ovarian carcinomas, and that it is therefore an important marker for the diagnosis and treatment of cancers, particularly breast and ovarian carcinomas. It may be a marker for the diagnosis of carcinomas (e.g. epidermal cancers)
  • the protein may be further characterised in that it has an apparent Mr of about 170 KDa.
  • the protein has been found to be expressed by the human breast cancer cell lines MCF-7, T-470 and ZR-75.
  • the invention also comprises an immunogenic fragment or analogue thereof of the protein.
  • Immunogenic fragments for example epitopes, particularly immunogenic epitopes, may be readily identified and analogues (mimotopes) made (Geysen, H.M. et al, 1987, Journal of Immunological Methods, 102: 259-274).
  • the invention also comprises an agent which binds specifically to the protein, fragment or analogue thereof.
  • an agent may for example comprise an antibody or an antigen binding fragment thereof.
  • Antibodies and methods of their production are well known (Harlow, E. and Lane, D., "Antibodies - A Laboratory Manual”, Cold Spring Harbor Laboratory, Cold Spring Harbor Press, New York, 1988).
  • the antibody may be a whole antibody or an antigen binding fragment thereof and may in general belong to any immunoglobulin class. Thus, for example, it may be an immunoglobulin M antibody or an immunoglobulin G antibody.
  • the antibody or fragment may be of animal, for example, mammalian origin and may be for example of murine, rat, sheep or human origin. It may be a natural antibody or a fragment thereof, or, if desired, a recombinant antibody fragment, i.e. an antibody or antibody fragment which has been produced using recombinant DNA techniques.
  • Particular recombinant antibodies or antibody fragments include, (1) those having an antigen binding site at least part of which is derived from a different antibody, for example those in which the hypervariable or complementarity determining regions of one antibody have been grafted into the variable framework regions of a second, different antibody (as described in, for example, EP 239400); (2) recombinant antibodies or fragments wherein non-Fv sequences have been substituted by non-Fv sequences from other, different antibodies (as described in, for example, EP 171469, 173494 and 194276); or (3) recombinant antibodies or fragments possessing substantially the structure of a natural immunoglobulin but wherein the hinge region has a different number of cysteine residues from that found in the natural immunoglobulin but wherein one or more cysteine residues in a surface pocket of the recombinant antibody or fragment is in the place of another amino acid residue present in the natural immunoglobulin (as described in, for example, PCT/GB 88/00730 and
  • the antibody or antibody fragment may be of polyclonal or monoclonal origin. It may be specific for at least one epitope.
  • Antigen binding antibody fragments include, for example, fragments derived by proteolytic cleavage of a whole antibody, such as F(ab')2,Fab' or Fab fragments, or fragments obtained by recombinant DNA techniques, for example Fv fragments (as described, for example, in International Patent Application No PCT/GB88/0747).
  • the antibodies according to the invention may be prepared using well-known immunological techniques.
  • any suitable host may be injected with the protein and the serum collected to yield the desired polyclonal antibody after appropriate purification and/or concentration (for example by affinity chromatography using the immobilised protein as the affinity medium).
  • splenocytes or lymphocytes may be recovered from the protein-injected host and immortalised using for example the method of Kohler et al. (1976, Eur. J. Immunol., 6: 511), the resulting cells being segregated to obtain a single genetic line producing monoclonal antibodies.
  • Antibody fragments may be produced using conventional techniques, for example, by enzymatic digestion with pepsin or papain. Where it is desired to produce recombinant antibodies according to the invention these may be produced using, for example, the methods described in EP 171469, EP 173494, EP 194276 and EP 239400.
  • Antibodies according to the invention may be labelled with a detectable label or may be conjugated with an effector molecule, for example a drug eg. an antibacterial agent or a toxin or an enzyme, using conventional procedures and the invention extends to such labelled antibodies or antibody conjugates.
  • an effector molecule for example a drug eg. an antibacterial agent or a toxin or an enzyme
  • the invention also comprises the protein, immunogenic fragment or analogue thereof, binding agent, antibody or antigen binding fragment thereof for use in a method of treatment or diagnosis of the human or animal body or when incorporated into a diagnostic kit therefor.
  • the invention also comprises the use of the protein, immunogenic fragment or analogue thereof, binding agent, antibody or antigen binding fragment thereof in the preparation of a medicament for treating cancer. Also provided is a method of manufacture of a medicament for treating cancer, characterised in the use of a protein, immunogenic fragment or analogue thereof, binding agent, antibody or antigen binding fragment thereof.
  • Medicaments may also comprise a pharmaceutically acceptable carrier, diluent or excipient (Remington's Pharmaceutical Sciences and US Pharmacopeia, 1984, Mack Publishing Company, Easton, PA, USA).
  • a diagnostic test method for the protein comprisiing the steps of :
  • a diagnostic test may for example be a sandwich ELISA test, or a dip-stick test (WO 88/08534).
  • a diagnostic test kit for performing the diagnostic test.
  • a diagnostic test kit may include instructions for its use in detecting the protein.
  • Also provided according to the present invention is the use of the protein, immunogenic fragment or analogue thereof, binding agent, antibody or antigen binding fragment thereof in the manufacture of a diagnostic test kit for the protein.
  • a method of treatment or diagnosis of the human or animal body comprising the use of the protein, immunogenic fragment or analogue thereof, binding agent, antibody or antigen binding fragment thereof according to the present invention.
  • a method of diagnosis may comprise the steps detailed above for a diagnostic test method.
  • a method of treatment may comprise administering to a patient the the protein, immunogenic fragment or analogue thereof, binding agent, antibody or antigen binding fragment thereof in a pharmacuetically acceptable dosage.
  • Pharmaceutically acceptable doses may be readily determined using standard dose-response assays.
  • the invention also comprises a nucleotide sequence encoding the protein; such a sequence may comprise at least the sequence of SEQ ID NO: 1 (pBl) or a partially modified form thereof.
  • the invention also comprises a DNA construct comprising such a coding sequence operably linked to at least one regulatory expressive signal such that the protein, or the fragment or analogue thereof may be expressed.
  • a construct may comprise, operatively linked in sequence in the 5' to 3' direction: i) a promoter region that directs the transcription of a gene;
  • Figure 1 shows the partial nucleotide sequence designated pBl (SEQ ID NO: 1) of a cDNA transcript of an mRNA species of 4.8Kb, and its two open reading frames ORF1 (SEQ ID NO: 2) and ORF2 (SEQ ID NO: 3);
  • Figure 2 shows MUC-B1 immunostaining on normal breast tissue (right) and breast carcinoma tissue (left) using a polyclonal anti-MUC-B 1 antiserum (rabbit) in a 1 :500 dilution.
  • Figure 3 shows MUC-B1 immunostaining on sections of a f ⁇ broadenoma using a polyclonal anti-MUC-B 1 antiserum (rabbit) in a 1 :500 dilution, magnification xlOO;
  • Figure 4 shows MUC-B1 immunostaining on a colon carcinoma section using a polyclonal anti-MUC-B 1 antiserum (rabbit) in a 1 :500 dilution.
  • Figure 5 shows MUC-B 1 immunostaining on a section of an ovarian carcinoma using a polyclonal anti-MUC-B 1 antiserum (rabbit) in a 1 :500 dilution.
  • Magnification xlOO shows MUC-B1 immunostaining on a colon carcinoma section using a polyclonal anti-MUC-B 1 antiserum (rabbit) in a 1 :500 dilution.
  • MUC-B 1 The following experiments detail the isolation and purification of the protein of the present invention (termed MUC-B 1) and the sequencing of it, identifying two open reading frames (ORFs), both of which are found to be expressed and the two expression products recognised by both polyclonal and monoclonal antibody specific against MUC- B 1.
  • Results show (see Results section, Tables 1 and 2) that anti-MUC-B 1 antibody detects substantial proportions of breast, ovarian and colon carcinomas.
  • MUC-B 1 and antibody specific against it is an extremely useful tool in the diagnosis of tumours, and thus also has the potential for use in treatment of same tumours.
  • mAb B-l A monoclonal antibody, designated mAb B-l has been established directed against the antigen present in Mz-Sto-1 and used in an immunoscreening procedure to isolate specific cDNA transcript from the following cDNA libraries :
  • pBl partial cDNA transcripts of a novel mRNA species of 4.8 kb were isolated.
  • pBl was isolated from library (a) and contains a insert 798 bp
  • pB 1/1.1 was isolated from the cDNA library (b) and contains an insert of 1191 bp. They share a completely identical sequence. Due to the internal homology structure within the nucleotide sequence, clone pB 1/1.1 could not be completely sequenced and hence did not provide additional sequence information. All sequence data revealed from the clone were identical to the sequence of pBl.
  • the coding sequence of the novel type of mRNA that has given rise our two clones is composed of multiple tandem repeats of 51-54 nucleotides in length. Characteristically, each of these nucleotide repeat motifs contains a cleavage site for the restriction enzyme Earl. Both clones have two possible ORF's at the 5' end, terminated by a stop codon in the 3' portion, followed by a 3' untranslated region of 264 bp and a poly (A) + tail. As shown in Figure 1, both ORF's encode very similar peptides with multiple tandem repeat motifs (TGQRRGS - e.g.
  • the pBl deduced peptide (from the ORFl used in clone pBl) comprises 171 amino acids. This peptide has a high content of alanine (15.2%), glycine (14.6%), serine (10.5%), threonine (5.9%) and proline (9.4%), a composition that is very similar to that of other mucins. Hydrophobicity index of the deduced Bl peptide was calculated using the method described by Kyte-Doolittle and shown to be very hydrophilic, because only 8 in 171 amino acids show positive values.
  • MUC-B 1 The amino acid composition, the repetitive structure in the cDNA - derived protein sequence, the high molecular weight and the presence of this protein in a mucin fraction of HT-29 MTX cells provides strong evidence that it is a hitherto unrecognised member of the human mucin family, and is referred to herein as MUC-B 1.
  • Northern blot analysis reveals that MUC-B 1, unlike other mucins, is encoded by a single mRNA transcript of 4.8Kb.
  • a 5' sequence of the mRNA was obtained by isolating additional cDNA clones and a cosmid representing the 3'-end of the MLP gene.
  • MUC-B 1 is not expressed by normal breast epithelium, whereas it is found in approximately 60% of specimens of breast carcinoma, indicating that MLP expression is an important marker for diagnosis, prognosis and monitoring of breast cancer, as it is not, unlike MUC1, expressed by normal breast epithelium.
  • MUC-B 1 is, moreover, not specifically filtered by the kidney, so tumours may be easily detected by testing for MUC-B 1 in blood.
  • Formalin-fixed, paraffin embedded tissue was used. 4mm sections were dewaxed and rehydrated. They were incubated with mAb-Bl cell culture supernatant diluted 1 :10 in Tris-saline buffer pH 7.8 for 18 hours at 4°C.
  • MCF-7, T-47D, MDA-MB-231 and HBL100 breast cell lines were obtained from the ATTCC. They were cultured in phenol-red free Dulbecco's modified Eagles 's medium supplemented with 10% foetal bovine serum and 2mM L-glutamine in a humidified atmosphere of 5% C0 2 /95% air at 37°C until 70-90% confluence.
  • cells were detached using Trypsin-EDTA and cultured on poly- D-lysine coated glass coverslips until 80% confluence.
  • immunoblotting cells were lysed using 1ml of TRI reagent (Sigma) per 10 cm 2 cells.
  • Peroxidase was localised using diaminobenzidine-hydrogen peroxide. Controls were omission of primary antibody and inclusion of pre-immune rabbit serum with each staining batch. For immunocytochemistry, cells were treated with acetone at 4 °C for 10 minutes prior to the above staining procedure.
  • Recombinant human MUC-B 1 was expressed in E. coli using the Thiofusion Expression System from Invitrogen, which generates thioredoxin fusion proteins. Native thioredoxin localises to an osmotically sensitive compartment between the cell wall and the palsma membrane in E. coli. Thiofusion proteins frequently do the same, which permits simple purification by osmotic shock, using hypotonic buffers.
  • Recombinant human MUC-B 1 protein was expressed as a thioredoxin fusion protein in E. coli using plasmid pTrxfus (Invitrogen BV, Leek, The Netherlands).
  • the pTrxfus plasmid was cut with the restriction endonucleases BamHI and Xbal.
  • Clone pB 1 (see figure 1) was used as a template for PCR with primers for MUC-B 1 5' / BamHI pTRX of SEQ ID NO: 4 (5' GGGGATCCGG CACGAGAGAA GAGGAAGCCG 3') and for MUC-B 1 3' / Xbal pTRX of SEQ ID NO: 5 (5' TCAGAATTCC GCACGAGAGA AGAGGAAGCC G 3').
  • PCR amplification was done in a 50 ⁇ l reaction volume containing 60 pmoles of each primer, 10 nmoles of each dNTP, 20 ng of cDNA template and 2U Taq polymerase (Gibco-BRL Life Technologies, Paisley, England) in the manufacturer's incubation buffer. Thirty cycles of amplification were performed, each composed of 45 seconds at 94 °C, 45 seconds at 63 °C and 1.5 minutes at 72 °C. The initial denaturation step was prolonged to 2 minutes and the final extension step to 5 minutes.
  • the expected 1115 bp PCR product was obtained and ligated into pCRl 1 (RTM, Invitrogen) Plasmid DNA was prepared, digested with BamHIIXbal and separated on an agarose gel. After electrophoresis the 550 bp fragment was cut from the gel, extracted using the Sephaglas Bandprep kit (Pharmacia, Uppsala, Sweden), and then ligated into the previously prepared pTrxfus vector (see above). The construct (MUC-B 1/Trxfus) was transformed into E. coli strain GI724 (Invitrogen) whereafter plasmid DNA was isolated and sequenced to confirm that the subcloning had been successful.
  • RTM Invitrogen
  • a single colony carrying MUC-B 1/Trxfus was innoculated into 3 ml RMG-Amp (1XM9 salts, 2% casamino acids.0.5% glucose, 1 mMMgCl 2 and 100 ⁇ g/ml ampicillin; protocol - Invitrogen) and grown overnight at 30°C. The next morning 100 ml induction medium (lxM9 salts.0.2% casamino acids, 0.5% glucose, lmM MgCl 2 and 100 ⁇ g/ml ampicillin) was inoculated with the 3ml culture and incubated at 30°C until the absorbance at 550 nm was -0.5.
  • Protein expression was induced by adding lOO ⁇ g/ml L-tryptophan followed by additional culturing for 4 hours at 37°C before the cells were harvested by centrifugation.
  • the cell pellet was resuspended in 2.5 ml TSB (200 mM NaCl, 100 mM Tris-HCl, lmM EDTA pH 7.0) with 0.1 mM PMSF and lysed by 3 cycles of sonication, rapid freezing and thawing. Centrifugation of the sample at 12,000g gave a clear supernatant containing 8 mg/ml soluble protein.
  • thioredoxin peptide As a control, 12 kDa thioredoxin peptide was expressed in E. coli carrying plasmid pTrxfus. Control peptide was purified by osmotic shock, a procedure that takes advantage of the ability of thioredoxin to localize to an osmotically sensitive compartment under the plasma membrane in E. coli (described above). Osmotic shock was carried out according to the manufacturer's protocol.
  • the fusion protein was released by osmotic shock treatment according to the manufacturer's instructions and the lysate purified further by FPLC (fluid pressure liquid chromatography) using a Mono Q 5/5 column (Pharmacia) connected to a FPLC system. Proteins were separated at a flow rate of 1 ml/minute using a 40 ml linear gradient of 0- 300 mM NaCl. The fusion protein was eluted between 120-150 mM NaCl. Mono Q fractions containing the thiofusion protein were pooled and free recombinant MUC-B 1 was generated by proteolytic cleavage using enterokinase (EKMax) as recommended by Invitrogen.
  • EKMax enterokinase
  • a total volume of 3 ml was digested in 1 ml batches, each containing 200 mg of protein and 60 units of enterokinase enzyme. Digestion was carried out overnight at 37 °C. Cleavage products were separated by re-chromatography, as described above. Free recombinant MUC-B 1 eluted between a NaCl concentration of 170-180 mM, and was homogeneous in purity, as judged by 15% (w/v) SDS-PAGE, stained with Coomassie by using the Bio-Rad protein assay kit. Typically, a 50 ml culture containing a total of 75-80 mg of cells produced 1 mg of homogeneous MUC-B 1.
  • NZW rabbit was injected sub-cutaneously at multiple sites with 100 ⁇ g of affinity purified MUC-B 1 protein in 500 ⁇ l of PBS plus 1 volume incomplete Freund's adjuvant. Three further identical imunizations were done at fortnightly intervals before the rabbit was bled and anti-sera assayed by Western blot.
  • Anti-human MUC-B 1 polyclonal anitbodies were prepared as described in Ghebrehiwet, B. et al. (1994, J. Exp. Med., 179: 1809).
  • Preparative scale protein expression was carried out according to the following protocol: 1. Innoculate 3 ml of RMG-Amp with 20 ⁇ l of a glycerol stock of E. coli G1724 harbouring the appropriate pTrxfus construct. Grow overnight at 30 °C in a shaking incubator.
  • the cell wall of E.coli can be disrupted (manufacturer's instructions) by transferring the cells from a high ionic strenth buffer (osmotic shock solution # 1) to a buffer of low ionic strength (osmotic shock solution # 2). This process releases the contents of the periplasmic space but leaves the rest of the cell intact. Since the periplasmic space contains relatively few native E. coli proteins, highly expressed thiofusion proteins which localise to the periplasmic space often account for more then 90% of the protein released by osmotic shock.
  • Expression is under the control of the P L promoter from bateriophage ⁇ , which is one of the most efficient promoters for bacterial expression.
  • the bacteriophage ⁇ cl repressor binds to the operator region upstrean of the P L promoter and controls the level of expression from this promoter. Expression of c/repressor is in turn under the control of the trp repressor.
  • E. coli carrying pTrxfus are grown in tryptophan-free medium, the cl repressor is expressed and binds to the P L promoter, preventing transcription. Expression is induced by adding tryptophan to the medium, which shuts down cl repressor synthesis, so allowing transcription from the P L promoter.
  • pTrxfus was propagated in E. coli strain G1698, which expresses the cl repressor.
  • the bateria were grown at 30°C in special tryptophan-free medium to prevent premature expression of the MUC-B 1 -Thiofusion protein. Higher temperatures might have caused uncontrolled low-level transcription through the P L promoter.
  • the thioredoxin fusion partner normally enhances the solubility of the recombinant protein, preventing the formation of inclusion bodies. However, if insouble protein is produced, solubility may be further enhanced by expression at low temperatures (18 - 30 °C) for longer periods of time. The optimal conditions are determined empirically by as series of preliminary experiments.
  • the protein extracts from MCF-7 and T-47D breast cancer cells were eletrophoresed on 5% Polyacrylamide gels with SDS using a minigel-eletrophoresis unit (BioRad) at 50 rnA (Sambrook, J. et al, 1990, Molecular cloning: A laboratory manual Cold Spring Harbor Labratory Press, Cold Spring Harbor, NY). Protein concentrations were determined using a standard Bradford assay. The gel was blotted to nitrocellulose according to standard pocedures.
  • Non malignant breast
  • Immunoblotting of homogenates of MCF-7 and T47D cells showed one band of 290 kDa in both and one band of 195 kDa in MCF-7 and 180 kDa in T-47D cells (not shown).
  • the adenosquamous carcinoma, MMT, 2 mucinous and one endometrioid carcinoma were negative.
  • CTGCAGCTCC AGTGCCAACC AGCAGGGGGC ACAGGACAGA GAAGAGGAAG CGGCCAAGGC 240

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Abstract

La présente invention porte sur un marqueur (protéine) utilisé dans le traitement et le diagnostic des cancers, ainsi que sur l'utilisation de cette protéine, d'un fragment immunogène ou d'un analogue de celle-ci, ainsi que sur des agents de liaison spécifiques contre celle-ci, sur l'utilisation de cette protéine dans la fabrication de médicaments et sur des kits de test et des procédés de test. L'invention porte également sur une séquence nucléotidique codant ladite protéine.
PCT/GB1998/001184 1997-04-23 1998-04-23 Marqueur utilise dans le diagnostic de cancers WO1998048014A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU70677/98A AU7067798A (en) 1997-04-23 1998-04-23 Cancer marker

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GB9708190.5 1997-04-23
GBGB9708190.5A GB9708190D0 (en) 1997-04-23 1997-04-23 Cancer marker

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WO1998048014A1 true WO1998048014A1 (fr) 1998-10-29

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017214186A1 (fr) 2016-06-09 2017-12-14 University Of Leicester Anticorps monoclonaux, compositions et méthodes de détection de la protéine de type mucine (mlp) en tant que biomarqueur du cancer de l'ovaire et du pancréas

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991009867A1 (fr) * 1989-12-22 1991-07-11 Imperial Cancer Research Technology Limited Nucleotides de mucine
DE19547324A1 (de) * 1995-12-19 1997-06-26 Behringwerke Ag Humanes Muzin (MUC8)

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991009867A1 (fr) * 1989-12-22 1991-07-11 Imperial Cancer Research Technology Limited Nucleotides de mucine
DE19547324A1 (de) * 1995-12-19 1997-06-26 Behringwerke Ag Humanes Muzin (MUC8)

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
WALTER A ET AL: "CLONING AND EXPRESSION OF A NOVEL MEMBER OF THE MUCIN FAMILY", IMMUNOBIOLOGY, vol. 186, October 1992 (1992-10-01), pages 72, XP002075060 *
WALTER AO AND DIPPOLD W: "Homo sapiens B1 mRNA for mucin-like antigen (accession number X84838)", EMBL SEQUENCE DATABASE, HEIDELBERG, GERMANY, 1 September 1995 (1995-09-01), XP002075061 *
WALTER AO ET AL: "H. sapiens B1 mRNA for mucin (accession number X83412)", EMBL SEQUENCE DATABASE, HEIDELBERG, GERMANY, 1 June 1995 (1995-06-01), XP002075059 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017214186A1 (fr) 2016-06-09 2017-12-14 University Of Leicester Anticorps monoclonaux, compositions et méthodes de détection de la protéine de type mucine (mlp) en tant que biomarqueur du cancer de l'ovaire et du pancréas
JP2019526227A (ja) * 2016-06-09 2019-09-19 ユニバーシティー オブ レスター 卵巣がんおよび膵臓がんのためのバイオマーカーとしてムチン様タンパク質（ｍｌｐ）を検出するためのモノクローナル抗体、組成物および方法
US10626184B2 (en) 2016-06-09 2020-04-21 Omeros Corporation Monoclonal antibodies, compositions and methods for detecting mucin-like protein (MLP) as a biomarker for ovarian and pancreatic cancer
AU2017277288B2 (en) * 2016-06-09 2024-05-16 Omeros Corporation Monoclonal antibodies, compositions and methods for detecting mucin -like protein (MLP) as a biomarker for ovarian and pancreatic cancer

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GB9708190D0 (en) 1997-06-11

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