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WO1990006323A2 - Proteines chimeriques incorporant une proteine de liaison de metaux - Google Patents

Proteines chimeriques incorporant une proteine de liaison de metaux Download PDF

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Publication number
WO1990006323A2
WO1990006323A2 PCT/US1989/005424 US8905424W WO9006323A2 WO 1990006323 A2 WO1990006323 A2 WO 1990006323A2 US 8905424 W US8905424 W US 8905424W WO 9006323 A2 WO9006323 A2 WO 9006323A2
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WO
WIPO (PCT)
Prior art keywords
metal
immunoglobulin
tumor
protein
labelled
Prior art date
Application number
PCT/US1989/005424
Other languages
English (en)
Other versions
WO1990006323A3 (fr
Inventor
Hubert J. P. Shoemaker
John Ghrayeb
Lee K. Sun
Original Assignee
Centocor, Inc.
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.)
Filing date
Publication date
Application filed by Centocor, Inc. filed Critical Centocor, Inc.
Publication of WO1990006323A2 publication Critical patent/WO1990006323A2/fr
Publication of WO1990006323A3 publication Critical patent/WO1990006323A3/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • A61K51/1045—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody against animal or human tumor cells or tumor cell determinants
    • A61K51/1063—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody against animal or human tumor cells or tumor cell determinants the tumor cell being from stomach or intestines
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/825—Metallothioneins
    • 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
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/3046—Stomach, Intestines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2121/00—Preparations for use in therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2123/00—Preparations for testing in vivo
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Definitions

  • the first is the direct labelling method by which a radiometal is bound to the protein molecule itself.
  • the second is the indirect labelling method by which a bifunctional agent is coupled to the protein and the radiometal is attached to the protein through the bifunctional agent.
  • This invention pertains to chimeric proteins which can be stably labelled with a metal
  • the chimeric proteins comprise a protein having an affinity for a biological target linked to a metal binding protein (or the metal binding domain of a protein) which is not normally associated with the protein (i.e. derived from another protein or peptide).
  • the linkage is a peptide linkage such that the two components form a contiguous polypeptide. Incorporation of the metal binding region into a protein allows a protein to be labelled with a metal.
  • the labelled chimeric proteins are useful as diagnostic and therapeutic agents, preferably radiodiagnostic and radio- therapeutic agents.
  • the chimeric proteins of this invention are produced by incorporating a metal binding protein (or domain thereof) into the amino acid sequence of the protein specific for the biological target by genetic engineering techniques. Proteins specific for a biological target can include biological receptors such as immunoglobulins and immunoglobulin fragments and ligands for
  • biological receptors such as hormones and growth factors.
  • this invention pertains to recombinant immunoglobulins having a metal binding protein incorporated into one or more of their constituent chains and to immunoglobulins labelled with radiometals through the metal binding protein.
  • the recombinant immunoglobulins can be stably labelled with a metal or radiometal for immunodiagnostics and immunotherapeutic procedures.
  • Figure 1 shows the construction of a plasmid
  • Figure 2 shows the construction of an expression vector containing the DNA encoding the variable region of the 17-1A tumor-specific antibody (17-1A V H ) and DNA encoding the human constant region of an immunoglobulin linked to the DNA encoding human metallothionein (hC ⁇ 4 /MT).
  • Figure 3 is a graph showing the binding of radioiodinated murine 17-1A to HT29 cells in the presence of a purified IgG of 17-1A(o) or G4K/MT(o).
  • Figure 4 is a gel filtration HPLC chromatogram of Tc-99m-IgG2a.
  • Figure 5 is a gel filtration HPLC chromatogram of Tc-99m-IgG4K/MT. Detailed Description
  • the chimeric proteins of this invention are proteins which have an affinity for a biological target, into which a metal binding domain has been incorporated.
  • the metal binding domain is one which is not normally associated with the protein and allows the chimeric protein to be stably labeled with a trace-metal, preferably a radiometal.
  • a metal is any metal capable of binding to a metal binding region which can either be detected in vivo or serve as a toxin to target cells.
  • Metal binding proteins are proteins having an affinity for metals such as metallothionein. Examples of
  • inventions are technetium-99m, In-111, Cu-67, Pd-109, Pd-103, Re-188, Au-198, Au-199, Ru-97, Hg-197,
  • the radiometals Tc-99m, In-111, Cu-67, and Pb-203 can be used in diagnostic applications and Cu-67, Pd-109, Re-188, and Au-199 can be used in therapeutic applications.
  • An example of a non-radioactive metal which can serve as a toxin is zinc.
  • the chimeric protein can be produced by genetic engineering techniques.
  • DNA encoding the protein is ligated to the DNA encoding the metal binding protein or a sufficient protion of the metal binding domain.
  • a sufficient portion of the metal binding domain is that portion necessary for
  • the DNA construct is designed so that the the resulting DNA construct is inserted into an expression vector which is
  • the chimeric protein is an immunoglobulin protein into which a metal binding domain has been incorporated.
  • the immunoglobulins can be specific for various applications.
  • the immunoglobulins can be specific for tumor- or proliferation-associated antigens. These include antigens associated with gastrointestinal, breast, ovarian, lung and renal cancer cells. Some specific examples are the antigens defined by antibodies 17-1A (gastrointestinal tumors), 0C125 (ovarian carcinoma), 0V-TL3 (ovarian carcinoma), 103D2 (breast) and 123. C3 (renal carcinoma). Others include 72.3 (colon), DF3, 115D8, RC38 (renal), G250 (renal) and 55-2A.
  • the immunoglobulins can be specific for a variety of markers of cardiovascular disease.
  • Some examples include RllDlO (myosin-specific) which can be used to detect and localize and evaluate myocardial infarction, 7E3, 10E5, S12 (platelet- specific) and T2G1, 59D8 and GC4 (fibrin- specific) which can be used to detect thromboses and atherosclerotic plaque.
  • the metal binding protein is incorporated into at least one of the constituent chains of the immunoglobulin, preferably a heavy chain.
  • This can be accomplished by preparing a DNA construct comprising, at minimum, a DNA segment which encodes (1) at least the functional portion of the variable region of an immunoglobulin chain l inke d to (2) a DNA segment encoding the metal binding protein or its functional domain.
  • the DNA encoding the metal binding protein is linked to DNA encoding at least a portion of the constant region.
  • the DNA construct is assembled in or inserted into a DNA expression vector by standard techniques.
  • Recipient cells capable of expressing the encoded product are then transfected with the DNA construct.
  • the recipient cells are also transfected with the DNA encoding the counterpart chain.
  • the transfected recipient cells are cultured and the expressed immunoglobulins or immunoglobulin chains are recovered.
  • Genes encoding the variable region of Ig light and heavy chains can be obtained from lymphoid cells which produce the antibodies specific for the desired target antigen.
  • lymphoid cells which produce the antibodies specific for the desired target antigen.
  • the hybridoma cell lines which produce antibodies against the specific tumor antigens provide a source of
  • immunoglobulin variable region genes against those antigens can be produced by challenging a rodent with a tumor cell or a tumor antigen containing cell component or fraction, forming fused hybrid cells between antibody producing cells and a myeloma cloning the hybrid and selecting clones which produce antibody against tumor-associated antigen.
  • Preferred constant regions of the immunoglobulins are of human origin. Human constant regions can be obtained from antibody producing cells by standard cloning techniques. Alternatively, because genes representing the two classes of light chains and the five classes of heavy chains have been cloned, constant regions of human origin are readily available from these clones.
  • Genes encoding the metal binding region can be obtained from a DNA clone of the metal binding region or can be synthesized using standard
  • the preferred metal binding protein is metallothionein; however, other metal binding proteins or oligo- or polypeptides such as lys-cys-thr-cys-cys-ala can also be used.
  • the metal binding protein is a human protein because this reduces immunogenicity.
  • the DNA construct encoding Immunoglobulin chain/metal binding protein and the counterpart chain can be assembled in two different expression vectors which can be used to cotransform a recipient cell.
  • each vector contains two selectable genes-one for selection in a bacterial system and one for selection in a eukaryotic system- each vector having a different pair of such genes. These vectors allow production and amplification of the DNA constructs in bacterial systems and
  • selectable genes for the bacterial system are the genes which confer ampicillin and the gene which couples chloramphenicol resistance.
  • Two selectable genes for selection of eukaryotic transfectants are preferred: (i) the xanthine-guanine phosphoribosyl- transferase gene (gpt), and (ii) the phosphotrans- ferase gene from Tn5 (designated neo). Selection with gpt is based on the ability of the enzyme encoded by this gene to use xanthine as a substrate for purine nucleotide synthesis; the analogous endogenous enzyme cannot.
  • a myeloma cell can synthesize, assemble and secrete immunoglobulins encoded by transfected Ig genes. Further, it possesses the mechanism for glycosylation of the immunoglobulin.
  • a particularly preferred recipient cell is the myeloma cell Sp2/0. This cell produces only immunoglobulin encoded by the transfected immunoglobulin genes.
  • Myeloma cells can be grown in culture or in the peritoneum of mice where secreted immunoglobulin can be obtained from ascites fluid. Other lymphoid cells such as B lymphocytes or hybridoma cells can serve as suitable recipient cells.
  • lymphoid cell with vectors containing DNA coding for the Immunoglobulin chain/metal binding protein hybrid.
  • a preferred way of Introducing DNA into lymphoid cells is by electroporation.
  • the cells and appropriate expression vectors are placed in an electroporation apparatus in media and
  • transfectants are selected in a growth medium.
  • Other techniques which can be used to introduce DNA into many cell types are calcium phosphate precipitation, diethylaminoethyl
  • DNA constructs in appropriate vectors can be expressed in non-animal cells such as bacteria.
  • the immunoglobulin chains When expressed in bacteria, the immunoglobulin chains become part of inclusion bodies. Thus, the chains must be isolated and purified and then assembled into immunoreactive, radiometal-binding immunoglobulin molecules.
  • Radiolabeled proteins can be used in immunoscintigraphy.
  • One important use is the imaging of tumors.
  • Antibody fragments are preferred for most immunoscintigraphic techniques.
  • Labeled Fab' fragments of tumor specific antibodies can be prepared and used to image primary or secondary tumors or myocardial infarctions.
  • the preferred radioisotope of immunoscintigraphy is technetium-99m which has a single photon energy of 140 keV, a half-life of about 6 hours, and is readily available from a 99 Mo - 99m Tc generator.
  • the radiolabeled Fab' fragment is introduced into a subject. After it is introduced into the subject, sufficient time is allowed for the labeled
  • Fab' fragment to accumulate at the site of the tumor or myocardial infarcts.
  • the subject is then scanned with a gamma camera to detect the gamma emission of the technetium-99m and to thereby obtain an image of the myocardial infarction or tumor.
  • a tumor or infarction can be localized and its size can be determined.
  • radiolabeled proteins can be used in radioimmuno therapy by selectively delivering radioisotopes to cells in vivo.
  • the preferred radioisotope for radioimmunotherapy is rhenium since it is a beta emitter which can kill target cells.
  • Non-radioactive metals which are toxic to the target cells can also be used for immunotherapy.
  • Tumor specific labeled antibodies are introduced into a subject wherein the labeled antibodies selectively seek out and destroy cancer cells.
  • the oligonucleotides were prepared using the phosphoramidite method on Applied Biosystems DNA Synthesizer model 380A.
  • the crude product was purified on a 20% polyacrylamide gel.
  • the purified DNA was used as a primer in DNA sequencing for use in cloning.
  • the purified material was phosphorylated and annealed to form double-stranded fragments.
  • DNA fragments were subcloned into M13mp18 RF vector. DNA sequences were determined by the dideoxy sequencing method using appropriate primers.
  • transfected cells were selected in growth medium containing 1 mg/ml of G418 for the expression of the light chain.
  • transfected cells were selected in growth medium containing 1 ug/ml of mycophenolic acid, 50 ug/ml of xanthine and 2.5 ug/ml of
  • a stably transfected cell line G4K/MT was established and analyzed.
  • Concentration of IgG production was estimated to be 5 ug/ml by using polystyrene beads coated with goat anti-human IgG (Fc) antibody and fluorescein- conjugated goat anti-human IgG (Fc) antibody.
  • Stable transfected cell lines were carried in DMEM containing G418 at 1 mg/ml , myc opheno l ic at
  • xanthine at 50 ug/ml
  • hypoxanthine at 2.5 ug/ml
  • ZnCl 2 at 0.14 ug/ml, supplemented with 15% fetal bovine serum.
  • Tissue culture supernatant was passed through an affinity column of Sepharose-bound goat
  • EDTA ethylenadiamine tetraacetic acid
  • 100uM ZnCl 2 ethylenadiamine tetraacetic acid
  • Murine IgG 17-1A was purified from ascites fluid by chromatography on staphylococcal protein
  • a monolayer of HT-29 human colorectal carcinoma cells were harvested with trypsin, washed and resuspended in growth medium. Cells were then seeded into 96-well microliter plates at
  • iodinated murine 17-1A and different concentrations of cold competing IgG in a final volumn of 100ul.
  • the cells were then washed three times with warm medium and cell-bound radioactivity was measured in a gamma counter.
  • Plasmid phC ⁇ 4 containing, the human C ⁇ 4 genomic DNA was used to derive the IgG/metallothionein (MT) fusion gene. Plasmid phC ⁇ 4 was partially digested with restriction endonuclease SacII and ligated with synthetic 24-mer oligonucleotide,
  • the SacII recognition sequence CCGCGG , is located at nucleotide 1238-1243 downstream from the Sail site of phC ⁇ 4 .
  • hC7 contains 8 extra amino acids in addition to the 57 amino acids derived from hMT.
  • variable gene of 17-lA was isolated and cloned into an expression vector containing genomic DNA encoding human 3 constant region to give
  • the cell line G4K/MT was chosen for further analysis.
  • Antibody concentration in the culture supernatant of G4K/MT was estimated to be 5 ug/ml as measured by particle concentration fluorescence immunoas say.
  • a stannous chloride/D - glucarate composition vial was prepared as described in PCT Application WO88/07382, the teachings of which are hereby incorporated by reference.
  • This stannous chloride/D-glucarate composition was reconstituted with 1ml (10mCi) Tc-99m pertechnetate. After 10 minutes, 0.10 ml of the contents of the stannous chloride/D-glucarate vial was transferred to 0.10 ml IgG2a (1 mg/ml) in a second vial.
  • the reac ti on mixture was assayed by thin layer chromatography (ITLC; 0.1M citrate buffer, pH 5).
  • Tc-99m incorporation for IgG2a ranged from 1.7% to 0% while IgG4K/MT ranged from 42.5% to 65.0% (see Table 1).
  • Tc-99m labelled products i.e., IgG2a at 37°C for 5 hours and IgG4K/MT at 37° for 5 hours
  • HPLC gel filtration high performance liquid chromatography
  • IgG4K/MT can be accomplished by warming the antibody to 37° in the presence of Tc-99m glucarate. Under the same conditions, the IgG2a (17-1A) is not labelled with Tc-99m.

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  • Animal Behavior & Ethology (AREA)
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  • Genetics & Genomics (AREA)
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Abstract

Protéines chimériques comprenant une protéine présentant une affinité pour une cible biologique liée par l'intermédiaire d'une liaison peptidique à une protéine de liaison de métaux ou à un domaine fonctionnel de cette protéine qui n'est pas associé normalement avec ladite protéine. Cette protéine peut être une immunoglobuline spécifique de tumeurs ou d'antigènes associés à la prolifération, ou une immunoglobuline spécifique de marqueurs de maladies cardiovasculaires. Les protéines chimériques ci-décrites sont utilisées par des techniques de diagnostic in vivo telles que l'immunoscintigraphie et également comme agent d'immunothérapie.
PCT/US1989/005424 1988-11-29 1989-11-29 Proteines chimeriques incorporant une proteine de liaison de metaux WO1990006323A2 (fr)

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US27733088A 1988-11-29 1988-11-29
US277,330 1988-11-29

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WO1990006323A2 true WO1990006323A2 (fr) 1990-06-14
WO1990006323A3 WO1990006323A3 (fr) 1990-07-12

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Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992013572A1 (fr) * 1991-02-08 1992-08-20 Diatech, Inc. POLYPEPTIDES MARQUES AU TECHNETIUM 99m POUR LES TECHNIQUES D'IMAGERIE
US5169764A (en) * 1990-08-08 1992-12-08 Regeneron Pharmaceuticals, Inc. Multitrophic and multifunctional chimeric neurotrophic factors, and nucleic acids and plasmids encoding the chimeras
US5449761A (en) * 1993-09-28 1995-09-12 Cytogen Corporation Metal-binding targeted polypeptide constructs
US5659041A (en) * 1993-07-19 1997-08-19 Resolution Pharmaceuticals, Inc. Hydrazino-type radionuclide chelators having an N3 S configuration
WO1997033916A1 (fr) * 1996-03-12 1997-09-18 Centro De Inmunologia Molecular (Cim) Anticorps monoclonaux ior c5 pour le diagnostic et le traitement de tumeurs colorectales
US5783170A (en) * 1991-11-27 1998-07-21 Diatide, Inc. Peptide-metal chelate conjugates
US5849261A (en) * 1991-02-08 1998-12-15 Diatide, Inc. Radiolabeled vasoactive intestinal peptides for diagnosis and therapy
US5866097A (en) * 1991-11-27 1999-02-02 Diatide, Inc. Technetium-99m labeled peptides for imaging
US6391590B1 (en) * 1991-10-21 2002-05-21 The Regents Of The University Of California Recombinant streptavidin-metallothionein chimeric protein having biological recognition specificity
WO2006118615A2 (fr) * 2004-12-16 2006-11-09 Brandeis University Marqueur clonable pour la purification et le marquage par microscopie electronique
US7238340B1 (en) 1991-11-27 2007-07-03 Cis Bio International Monoamine, diamide, thiol-containing metal chelating agents
WO2008053229A1 (fr) * 2006-11-02 2008-05-08 Iti Scotland Limited Système de reconnaissance magnétique
WO2009133203A2 (fr) * 2008-05-02 2009-11-05 Iti Scotland Limited Imagerie in vivo
WO2018187031A1 (fr) * 2017-04-05 2018-10-11 Archer Daniels Midland Company Nouveau catalyseur d'estérification et ses utilisations

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4732864A (en) * 1983-10-06 1988-03-22 E. I. Du Pont De Nemours And Company Trace-labeled conjugates of metallothionein and target-seeking biologically active molecules
GB8422238D0 (en) * 1984-09-03 1984-10-10 Neuberger M S Chimeric proteins

Cited By (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5169764A (en) * 1990-08-08 1992-12-08 Regeneron Pharmaceuticals, Inc. Multitrophic and multifunctional chimeric neurotrophic factors, and nucleic acids and plasmids encoding the chimeras
WO1992013572A1 (fr) * 1991-02-08 1992-08-20 Diatech, Inc. POLYPEPTIDES MARQUES AU TECHNETIUM 99m POUR LES TECHNIQUES D'IMAGERIE
US5849261A (en) * 1991-02-08 1998-12-15 Diatide, Inc. Radiolabeled vasoactive intestinal peptides for diagnosis and therapy
US6391590B1 (en) * 1991-10-21 2002-05-21 The Regents Of The University Of California Recombinant streptavidin-metallothionein chimeric protein having biological recognition specificity
US5866097A (en) * 1991-11-27 1999-02-02 Diatide, Inc. Technetium-99m labeled peptides for imaging
US7238340B1 (en) 1991-11-27 2007-07-03 Cis Bio International Monoamine, diamide, thiol-containing metal chelating agents
US5985241A (en) * 1991-11-27 1999-11-16 Diatide, Inc. Peptide-metal chelate conjugate complexes
US5981477A (en) * 1991-11-27 1999-11-09 Diatide, Inc. Peptide-metal chelate conjugates
US5783170A (en) * 1991-11-27 1998-07-21 Diatide, Inc. Peptide-metal chelate conjugates
US5659041A (en) * 1993-07-19 1997-08-19 Resolution Pharmaceuticals, Inc. Hydrazino-type radionuclide chelators having an N3 S configuration
US5609847A (en) * 1993-09-28 1997-03-11 Cytogen Corporation Treatment methods using metal-binding targeted polypeptide constructs
US5593656A (en) * 1993-09-28 1997-01-14 Cytogen Corporation Metal-binding targeted polypeptide constructs
US5578288A (en) * 1993-09-28 1996-11-26 Cytogen Corporation Metal-binding targeted polypeptide constructs
US5449761A (en) * 1993-09-28 1995-09-12 Cytogen Corporation Metal-binding targeted polypeptide constructs
WO1997033916A1 (fr) * 1996-03-12 1997-09-18 Centro De Inmunologia Molecular (Cim) Anticorps monoclonaux ior c5 pour le diagnostic et le traitement de tumeurs colorectales
WO2006118615A2 (fr) * 2004-12-16 2006-11-09 Brandeis University Marqueur clonable pour la purification et le marquage par microscopie electronique
WO2006118615A3 (fr) * 2004-12-16 2007-03-08 Univ Brandeis Marqueur clonable pour la purification et le marquage par microscopie electronique
WO2008053229A1 (fr) * 2006-11-02 2008-05-08 Iti Scotland Limited Système de reconnaissance magnétique
EP2428799A1 (fr) * 2006-11-02 2012-03-14 ITI Scotland Limited Système de reconnaissance magnétique
WO2009133203A2 (fr) * 2008-05-02 2009-11-05 Iti Scotland Limited Imagerie in vivo
WO2009133203A3 (fr) * 2008-05-02 2010-08-05 Iti Scotland Limited Imagerie in vivo
WO2018187031A1 (fr) * 2017-04-05 2018-10-11 Archer Daniels Midland Company Nouveau catalyseur d'estérification et ses utilisations

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