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WO1993014098A1 - Antigene reconnu chez les patients avec le syndrome myasthenique de lambert. eaton (lems) associe a l'anticorps, adn codant celui-ci et son utilisation - Google Patents

Antigene reconnu chez les patients avec le syndrome myasthenique de lambert. eaton (lems) associe a l'anticorps, adn codant celui-ci et son utilisation Download PDF

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Publication number
WO1993014098A1
WO1993014098A1 PCT/US1993/000227 US9300227W WO9314098A1 WO 1993014098 A1 WO1993014098 A1 WO 1993014098A1 US 9300227 W US9300227 W US 9300227W WO 9314098 A1 WO9314098 A1 WO 9314098A1
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antibody
myasthenic
nucleic acid
antigenic polypeptide
polypeptide
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PCT/US1993/000227
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English (en)
Inventor
Henry M. Furneaux
Jerome B. Posner
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Sloan-Kettering Institute For Cancer Research
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Publication of WO1993014098A1 publication Critical patent/WO1993014098A1/fr

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Classifications

    • 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
    • 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
    • A61K51/1027—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 receptors, cell-surface antigens or cell-surface determinants
    • 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
    • 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

Definitions

  • Lambert-Eaton myasthenic syndrome is a paraneoplastic disorder of the neuromuscular junction characterized by a defect in the presynaptic quantal release of acetylcholine [1]. At least seventy percent of the patients with LEMS have small cell carcinomas of the lung [2], LEMS patients develop an immune response against component (s) of a voltage-gated CA2+ channel (VGCC) expressed in small cell carcinomas of the lung [3]. It has been suggested that this immune response cross reacts with Ca++ channels in the presynaptic neuromuscular junction and is responsible for the neurological dysfunction.
  • VGCC voltage-gated CA2+ channel
  • Paraneoplastic neurological syndromes are disorders of nervous system function that occur in association with cancer but not as a direct effect of the tumor or metastasis [5, 6]. An immune basis has been suggested for several of these disorders [7, 8, 9, 10], but LEMS is the prototype for which direct evidence of a pathogenic antibody(s) has been demonstrated by passive transfer of the disease to animals [11]. Clinical symptoms of LEMS include: proximal muscle weakness, a reduction or absence of tendon reflexes, and cholinergic dysautonomia [2]. Electophysiological studies show low amplitude of the compound muscle action potential that increases progressively during high frequency repetitive stimulation [1].
  • the VGCC consists of 5 subunits ( ⁇ 1, ⁇ 2, ⁇ , ⁇ , ⁇ ) .
  • the ⁇ 1 subunit is the central ion-channel component [16].
  • the function of the ⁇ 2- ⁇ subunits are not clear but may contain the ⁇ -conotoxin binding site [17].
  • the ⁇ subunit interacts with the ⁇ 1 subunit and may function in a regulatory role [15].
  • the ⁇ subunit has been shown to accelerate both activation and inactivation of the calcium channel current and to increase the number of calcium antagonist binding sites upon transfection into LCa.11 cells and so appears to have an important regulatory function [18].
  • the role of the ⁇ -conotoxin-sensitive-VGCC in the etiology of LEMS is uncertain as patients with SCLC and no detectable neurological dysfunction also harbor antibodies against ⁇ -conotoxin labelled VGCCs [13].
  • LEMS Lambert-Easton Myasthenic Syndrome
  • a method of detecting an antibody associated with Lambert- Easton Myasthenic Syndrome comprises contacting a suitable sample with a purified antigenic polypeptide labelled with a detectable marker under conditions so as to form a complex between the purified myasthenic antigenic polypeptide and the antibody, detecting the presence of any complex so formed, thereby detecting an antibody associated with Lambert-Easton Myasthenic Syndrome (LEMS).
  • LEMS Lambert- Easton Myasthenic Syndrome
  • Also provided by this invention is a method of determining whether a patient exhibiting neurological symptoms harbors antigenic polypeptides associated with Lambert-Easton Myasthenic Syndrome (LEMS), which comprises contacting a suitable tissue or cell sample from the patient with a monoclonal antibody directed against a myasthenic antigen, the monoclonal antibody being labeled with a detectable marker, under suitable conditions so as to form a complex between the antibody and the antigen, detecting the presence of any complex so formed, the presence of complex being a positive determination that the patient harbors antigens associated with Lambert-Easton Myasthenic Syndrome (LEMS).
  • LEMS Lambert-Easton Myasthenic Syndrome
  • Figure 1 Western blot analysis of mysB fusion protein.
  • E. coli protein (1-6) and mysB fusion protein (7-12) reacted with a normal human serum (1-3, 7-9) and LEMS serum (4-6, 10-12). Both sera were tested at three dilutions: 1/10,000, lanes 1, 4, 7, 10; 1/5000, lanes 2, 5, 8, 11; 1/1000, lanes 3, 6, 9, 12.
  • Figure 2 Nucleotide sequence of p mys A cDNA clone. Also sequence I.D. no. 1.
  • Figure 3 Nucleotide sequence of p mys B cDNA clone. Also sequence I.D. no. 2.
  • Figure 4 Predicted amino acid sequence of mysB cDNA clone and its homology to the ⁇ -subunit of rabbit calcium channel (RABCAC). Sequence analysis was done by the dideoxy termination method [20] using Sequenase 2.0 (United State Biochemical). Double stranded DNA was prepared using the Qiagen plasmid midi-prep system and seguenced on both strands. Single stranded DNA was prepared using a modified single strand rescue protocol (Stratagene). Internal oligonucleotide primer, SK and KS primers were used. (Sequence I.D. Nos. 3 and 4, respectively). Detailed Description of the Invention
  • myasthenic antigenic polypeptide encompasses any amino acid sequence, polypeptide or protein having the biological activity of a myasthenic antigenic protein, i.e., a protein which may specifically form a complex with an antibody which is characteristic of Lambert- Eaton Myasthenic Syndrome (LEMS).
  • LEMS Lambert- Eaton Myasthenic Syndrome
  • This antigen is also called myasthenic syndrome antigen.
  • the antibodies which may specifically form a complex with myasthenic antigenic polypeptide is characteristically found in patients with Lambert-Eaton Myasthenic Syndrome, a disorder of the brain found in association with neoplasms of lung.
  • the isolated nucleic acid sequences described hereinabove are DNA. In other embodiments of this invention, the isolated nucleic acid sequences described hereinabove are cDNA, or RNA. In the preferred embodiment of this invention, the isolated nucleic acid sequences are cDNA sequences as shown in Figures 2 and 3. cDNA clones encoding two antigenic polypeptides of the subject invention were deposited with the American Type Culture Collection (ATCC), 12301 Parklawn Drive, Rockville, Maryland 20852, U.S.A. under the provisions of the Budapest Treaty for the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. The two clones, designated mys A and mys B, were accorded ATCC designation numbers 75147 and 75137, respectively. Deposited mys B has the sequence depicted in Figure 3 from nucleotide 1 to nucleotide 424.
  • DNA sequences of the subject invention also include DNA sequences coding for polypeptide analogs, fragments or derivatives of antigenic polypeptides which differ from naturally-occurring forms in terms of the identity or location of one or more amino acid residues (deletion analogs containing less than all of the residues specified for the antigen, substitution analogs wherein one or more residues specified are replaced by other residues and addition analogs where in one or more amino acid residues is added to a terminal or medial portion of the polypeptide) and which share some or all the properties of naturally- occurring forms.
  • sequences include: the incorporation of codons "preferred" for expression by selected non-mammalian hosts; the provision of sites for cleavage by restriction endonuclease enzymes; and the provision of additional initial, terminal or intermediate DNA sequences that facilitate construction of readily expressed vectors.
  • the DNA sequences of this invention encode the full length unprocessed amino acid sequence as well as DNA sequences encoding the processed form of the antigen (a cDNA clone). Methods of making these analogs are well known to those of skill in the art, utilizing the information disclosed herein.
  • DNA sequences described and claimed herein are useful for the information which they provide concerning the amino acid sequence of the polypeptide and as products for the large scale synthesis of the polypeptide by a variety of recombinant techniques. These sequences are useful for generating new viral and circular plasmid vectors, transformed and transfected procaryotic and eucaryotic host cells (including bacterial and yeast cells and mammaliain cells grown in culture), and new and useful methods for cultured growth of such host cells capable of expression of the polypeptide and related products.
  • nucleic acid sequences are useful for the development of probes to study the immune reaction.
  • detectable markers such as a radioisotope, enzyme or dye
  • the nucleic acids may be employed in hybridization, histological or serological assays.
  • a hybridization probe is useful for locating the human gene position in a chromosomal map.
  • Nucleic acid sequences detectably labeled also are provided by this invention. Methods to label nucleic acids are well known to those skilled in the art.
  • vectors which comprise the isolated nucleic acid molecules described hereinabove also are provided.
  • Suitable vectors comprise, but are not limited to, a plasmid or a virus. These vectors may be transfected into a suitable host cell to form a host cell vector system for the production of a polypeptide having the biological activity of the myasthenic antigenic polypeptide.
  • This invention further provides an isolated DNA or cDNA molecule described hereinabove wherein the host cell is selected from the group consisting of bacteria cells (such as E. coli), yeast cells , fungi cells, insect cells and animal cells.
  • bacteria cells such as E. coli
  • yeast cells fungi cells
  • insect cells such as E. coli
  • animal cells include, but are not limited to Vero cells, HeLa cells, Cos cells , CV1 cells and primary mouse cells.
  • this invention is a method for producing a polypeptide having the biological activity of the myasthenic antigenic polypeptide comprising the steps of: a) culturing the host vector system described hereinabove under suitable conditions permitting production of the polypeptide, and b) recovering the polypeptide produced.
  • This invention also provides the polypeptides produced by this method.
  • references to specific nucleotides are to nucleotides present on the coding strand of the nucleic acid.
  • the following standard abbreviations are used throughout the specification to indicate specific nucleotides:
  • This invention also encompasses DNAs and cDNAs which encode amino acid sequences which differ from those of the myasthenic antigenic polypeptide, but which should not produce phenotypic changes.
  • this invention also encompasses DNAs and cDNAs which hybridize to the DNA and cDNA of the subject invention. Hybridization methods are well known to those of skill in the art.
  • This invention also encompasses cDNA and DNA molecules characterized by changes in non-coding regions that do not alter the phenotype of the polypeptide produced.
  • purified, myasthenic antigenic polypeptide shall mean an isolated naturally-occurring myasthenic polypeptide or protein (purified from nature or manufactured such that the primary, secondary and tertiary conformation, and the glycosylation pattern are identical to naturally-occurring material) as well as non-naturally occurring polypeptides having a primary structural conformation (i.e., continuous sequence of amino acid residues) and glycosylation sufficiently duplicative of that of naturally-occurring myasthenic antigenic polypeptide or protein to allow possession the biological activity of naturally-occurring myasthenic antigenic polypeptides and proteins.
  • Such polypeptides include derivatives and analogs.
  • the purified myasthenic antigenic polypeptide may be labeled with a detectable marker.
  • detectable markers include, but are not limited to detectable markers selected from the group consisting of radioisotopes, dyes, enzymes and biotin. These are useful in immunological assays to detect antibody to LEMS in a patient.
  • a human fetal brain expression library with the serum of a patient with LEMS resulting in the isolation of two independent but related cDNA clones encoding antigens designated myasthenic syndrome antigen A and B (mysA, mysB). Fusion proteins derived from these clones were not recognized by normal human sera or by sera from patients with SCLC. The amino acid sequence of these clones show high homology to the ⁇ subunit of the rabbit skeletal muscle Ca++ channel [4].
  • This invention further provides a monoclonal antibody directed to an epitope on the myasthenic antigenic polypeptide.
  • the monoclonal antibody is a mouse monoclonal antibody.
  • the monoclonal antibody is a human monoclonal antibody.
  • the monoclonal antibodies of the subject invention are useful for the immunogical detection of LEMS antigen in a patient sample.
  • mice For the isolation of mouse monoclonal antibodies, eight week old mice may be injected interperitoneally with about 50 micrograms of a synthetic, purified myasthenic antigenic polypeptide, (prepared as described above) in complete Freud's adjuvant 1:1 volume. Mice are boosted, at monthly intervals, with the polypeptide, mixed with incomplete Freund's adjuvant, and bled through the tail vein. On days 4, 3, and 2 prior to fusion, mice are boosted intravenously with 50 micrograms of the polypeptide in saline. Splenocytes are fused with non-secreting myeloma cells according to procedures which have been described and are known to those of skill in the art to which this invention pertains. Some time later, approximately two weeks later, hybridoma supernatant is screened for binding activity against the myasthenic antigenic polypeptide as described below.
  • Positive clones are then isolated and propagated. Isolates of human monoclonal antibodies will be similar except ⁇ cells will be isolated from patients and transformed with EBV. ⁇ cells will then be fused with non-secreting myeloma cells according to procedures which have been described and are known to those of skill in the art to which this invention pertains. Some time later, approximately two weeks later, hybridoma supernatant are then be screened for binding activity against the myasthenic antigenic polypeptide as described hereinafter. Positive clones are isolated and propagated.
  • compositions comprising the purified, myasthenic antigenic polypeptides described hereinabove alone, or conjugated to any one of the following, a detectable marker, a therapeutic agent, or an imaging agent, as described hereinabove and a pharmaceutically acceptable carrier.
  • pharmaceutical compositions comprising the monoclonal antibody described hereinabove alone, or conjugated to any one of the following, a detectable marker, a therapeutic agent, or an imaging agent.
  • the term "pharmaceutically acceptable carrier” encompasses any of the standard pharmaceutical carriers, such as phosphate buffered saline solution, water, emulsions, such as a oil/water emulsions, and various types of wetting agents.
  • the pharmacuetical compositions of this invention are useful in imaging, therapy or in diagnostic assays.
  • a method of detecting an antibody associated with Lambert- Easton Myasthenic Syndrome (LEMS) also is provided by this invention. This method comprises contacting a suitable sample with a purified, myasthenic antigenic polypeptide described hereinabove under conditions so as to form a complex between the purified myasthenic antigenic polypeptide and the antibody, detecting the presence of any complex so formed, thereby detecting an antibody associated with paraneoplastic sensory neuronopathy.
  • Suitable samples include any sample suspected containing an antibody associated with Lambert-Eaton, such as cerberal spinal fluid and serum.
  • the synthetic, purified myasthenic antigenic polypeptide is labeled with a detectable marker selected from the group consisting of radioisotope, dye, enzyme and biotin.
  • a detectable marker selected from the group consisting of radioisotope, dye, enzyme and biotin.
  • suitable radioisotopes include, but are not limited to, 32 P, 35 S, and 131 I.
  • Also provided by this invention is a method of determining whether a patient exhibiting neurological symptoms harbors antibodies associated with Lambert-Easton Myasthenic
  • LEMS Lambert-Easton Myasthenic Syndrome
  • the monoclonal antibody is labeled with a detectable marker.
  • detectable markers include, but are not limited to a detectable marker selected from the group consisting of a radioisotope, dye, enzyme and biotin. Suitable radioisotopes have been described hereinabove. A method of determining whether a patient exhibiting neurological symptoms harbors a tumor expressing myasthenic antigen is provided by this invention.
  • This method comprises contacting a suitable sample from the patient with the antibody of this invention under suitable conditions so as to form a complex between the antibody and the myasthenic antigen, detecting the presence of any complex so formed, the presence of complex indicating that the patient harbors a tumor expressing myasthenic antigenic polypeptide.
  • the antibody is labelled with a detectable marker, e.g., a radioisotope, dye, enzyme or biotin.
  • This invention further provides a method of inhibiting the proliferation of neoplastic cells in a patient having LEMS which comprises administering to the patient an amount of the antibody described hereinabove, effective to inhibit the proliferation of neoplastic cells.
  • the therapeutic agent may include, but is not limited to a radioisotope, toxin, toxoid or chemotherapeutic agent.
  • a method of imaging neoplastic cells in a patient, wherein the neoplastic cells are associated with LEMS comprises administering to the patient an amount of an antibody described hereiabove, in an amount sufficient to form a detectable complex between the antibody and the myasthenic antigen associated with LEMS, imaging any complex so formed, and thereby imaging neoplastic cells in the patient.
  • the imaging agent may be, but is not limited to a radioisotope. Suitable radioisotopes are well known to those of skill in the art.
  • administering means a method of administering to the patient.
  • Such methods include, but are not limited to administration orally, intravenously, or parenterally. Administration of the agent may be effected continuously or intermittently, such that the amount of the therapeutic agent in the patient is effective to inhibit proliferation of neoplastic cells.
  • suitable therapeutic agents include radioisotopes, toxins, toxoids, and chemotherapeutic agents.
  • a ⁇ ZAP II fetal brain library (Stratagene) was screened at a density of 5 x 10 4 pfu per 150 mm plate of E. coli XL1- Blue. After a 6 hour incubation at 37°C plates were overlaid with filters soaked in 10 mM IPTG and incubated for 12 hours at 37°C.
  • FIG. 1 shows the Western Blot analysis of pMys B.Cells were isolated by centrifugation and lysed by resuspension in 2% SDS, 50 mM HEPES pH 5.5. Lysates (200ug) were resolved by 10% SDS/polyacrylamide gel electrophoresis and transferred to nitrocellulose [19].
  • nitrocellulose was-blocked with 5% Blotto and incubated with the indicated amount of serum for 2 hours at room temperature, washed with TBST, incubated with 1-125 Protein A as described above, dried and exposed to XAR5 film at -70°C.
  • Figure 4 shows the predicted amino acid sequence of mysB cDNA clone and its homology to the ⁇ -subunit of rabbit calcium channel (RABCAC). Sequence analysis was done by the dideoxy termination method [20] using Sequenase 2.0 (United Stated Biochemical). Double stranded DNA was prepared using the Qiagen plasmid midi-prep system and sequenced on both strands. Single stranded DNA was prepared using a modified single strand rescue protocol (Stratagene). Internal oligonucleotide primers, SK and KS primers were used.
  • Neuronal antinuclear antibody in sensory neuronopathy from lung cancer Neurology. 35: 538-543, 1985.
  • Tanabe, T. Takeshima, H., Mikami, A. et al.
  • GCCACTTCAA GTCTGCCTCT TAGCCCCACC CTAGCCTCTA ATTCACAGGG TTCTCAAGGT 1560

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Abstract

L'invention concerne une séquence d'acide nucléique isolée codant le polypeptide antigénique paranéoplasique myasthénique. Cette invention concerne également un polypeptide antigénique myasthénique purifié et des compositions contenant celui-ci. L'invention concerne en outre un anticorps monoclonal dirigé contre un épitope sur le poplypeptide antigénique paranéoplasique myasthénique, ainsi que des compositions contenant cet anticorps monoclonal. Cette invention décrit aussi des procédés de diagnostic et de traitement utilisant les compositions décrites ci-dessus.
PCT/US1993/000227 1992-01-10 1993-01-11 Antigene reconnu chez les patients avec le syndrome myasthenique de lambert. eaton (lems) associe a l'anticorps, adn codant celui-ci et son utilisation WO1993014098A1 (fr)

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US07/820,312 1992-01-10

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Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5618720A (en) * 1988-04-04 1997-04-08 Sibia Neurosciences, Inc. Cells expressing calcium channel α2 subunit-encoding DNA, optionally with a reporter gene for screening assays
US5686241A (en) * 1988-04-04 1997-11-11 Sibia Neurosciences, Inc. Probes and assays for calcium channel α2 subunit-encoding nucleic acids
US5726035A (en) * 1990-02-20 1998-03-10 Sibia Neurosciences, Inc. Recombinant production of mammalian calcium channel gamma subunits
US5874236A (en) * 1988-04-04 1999-02-23 Sibia Neurosciences. Inc. DNA encoding human calcium channel α-1A, β1, β-2, and β-4 subunits, and assays using cells that express the subunits
WO1999005172A3 (fr) * 1995-01-24 1999-04-15 Shoukat Dedhar Produits pharmaceutiques permettant de moduler la capacite de reponse aux hormones
US6096514A (en) * 1988-04-04 2000-08-01 Sibia Neurosciences, Inc. Human calcium channel compositions and methods
US6387696B1 (en) * 1988-04-04 2002-05-14 Merck & Co., Inc. Human calcium channel compositions and methods
US6518397B1 (en) 1997-07-24 2003-02-11 Shoukat Dedhar Pharmaceuticals for modulating hormone responsiveness
US6528630B1 (en) 1997-12-03 2003-03-04 Merck & Co., Inc. Calcium channel compositions and methods
US6653097B1 (en) * 1991-08-15 2003-11-25 Merck & Co., Inc. Human calcium channel compositions and methods

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989009834A1 (fr) * 1988-04-04 1989-10-19 The Salk Institute Biotechnology/Industrial Associ Compositions a canal de calcium et procedes associes
WO1991013077A1 (fr) * 1990-02-20 1991-09-05 The Salk Institute Biotechnology/Industrial Associates, Inc. Procedes et compositions de canaux de calcium

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989009834A1 (fr) * 1988-04-04 1989-10-19 The Salk Institute Biotechnology/Industrial Associ Compositions a canal de calcium et procedes associes
WO1991013077A1 (fr) * 1990-02-20 1991-09-05 The Salk Institute Biotechnology/Industrial Associates, Inc. Procedes et compositions de canaux de calcium

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
ANNALS OF NEUROLOGY, Volume 29, No. 3, issued March 1991, K. LEYS et al., "Calcium Channel Autoantibodies in the Lambert-Eaton Myasthenic Syndrome", pp. 307-314. *
CANCER RESEARCH, Volume 50, No. 13, issued 01 July 1990, E. SHER et al., "Voltage-Operated Calcium Channels in Small Cell Lung Carcinoma Cell Lines: Pharmacological, Functional, and Immunological Properties", pp. 3892-3896. *
FEBS LETTERS, Volume 291, No. 2, issued October 1991, M. PRAGNELL et al., "Cloning and Tissue-Specific Expression of the Brain Calcium Channel Beta-Subunit", pp. 253-258. *
MAYO CLINIC PROCEEDINGS, Volume 64, issued December 1989, V.A. LENNON et al., "Autoantibodies Bind Solubilized Calcium Channel-w-Conotoxin Complexes from Small Cell Lung Carcinoma: A Diagnostic Aid for Lambert-Eaton Myasthenic Syndrome", pp. 1498-1504. *
METHODS IN ENZYMOLOGY, Volume 152, issued 1987, R.C. MIERENDORF et al., "Gene Isolation by Screening Lambda gt11 Libraries with Antibodies", pp. 458-469. *
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES USA, Volume 89, issued April 1992, C. LEVEQUE et al., "The Synaptic Vesicle Protein Synaptotagmin Associates with Calcium Channels and is a Putative Lambert-Eaton Myasthenic Syndrome Antigen", pp. 3625-3629. *
THE LANCET, Volume 2, issued 16 September 1989, E. SHER et al., "Specificity of Calcium Channel Autoantibodies in Lambert-Eaton Myasthenic Syndrome", pp. 640-643. *

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5618720A (en) * 1988-04-04 1997-04-08 Sibia Neurosciences, Inc. Cells expressing calcium channel α2 subunit-encoding DNA, optionally with a reporter gene for screening assays
US5686241A (en) * 1988-04-04 1997-11-11 Sibia Neurosciences, Inc. Probes and assays for calcium channel α2 subunit-encoding nucleic acids
US5874236A (en) * 1988-04-04 1999-02-23 Sibia Neurosciences. Inc. DNA encoding human calcium channel α-1A, β1, β-2, and β-4 subunits, and assays using cells that express the subunits
US6013474A (en) * 1988-04-04 2000-01-11 Sibia Neurosciences, Inc. Calcium channel compositions and methods
US6096514A (en) * 1988-04-04 2000-08-01 Sibia Neurosciences, Inc. Human calcium channel compositions and methods
US6387696B1 (en) * 1988-04-04 2002-05-14 Merck & Co., Inc. Human calcium channel compositions and methods
US7063950B1 (en) * 1988-04-04 2006-06-20 Harpold Michael M Nucleic acids encoding human calcium channel and methods of use thereof
US5726035A (en) * 1990-02-20 1998-03-10 Sibia Neurosciences, Inc. Recombinant production of mammalian calcium channel gamma subunits
US6653097B1 (en) * 1991-08-15 2003-11-25 Merck & Co., Inc. Human calcium channel compositions and methods
WO1999005172A3 (fr) * 1995-01-24 1999-04-15 Shoukat Dedhar Produits pharmaceutiques permettant de moduler la capacite de reponse aux hormones
US6518397B1 (en) 1997-07-24 2003-02-11 Shoukat Dedhar Pharmaceuticals for modulating hormone responsiveness
US6528630B1 (en) 1997-12-03 2003-03-04 Merck & Co., Inc. Calcium channel compositions and methods

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