+

WO1986006487A1 - Methode de determination de mimotopes - Google Patents

Methode de determination de mimotopes Download PDF

Info

Publication number
WO1986006487A1
WO1986006487A1 PCT/AU1986/000110 AU8600110W WO8606487A1 WO 1986006487 A1 WO1986006487 A1 WO 1986006487A1 AU 8600110 W AU8600110 W AU 8600110W WO 8606487 A1 WO8606487 A1 WO 8606487A1
Authority
WO
WIPO (PCT)
Prior art keywords
monomers
catamers
catamer
receptor
solid support
Prior art date
Application number
PCT/AU1986/000110
Other languages
English (en)
Inventor
Hendrik Mario Geysen
Original Assignee
Commonwealth Serum Laboratories Commission
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 Commonwealth Serum Laboratories Commission filed Critical Commonwealth Serum Laboratories Commission
Priority to AT86902772T priority Critical patent/ATE78097T1/de
Priority to DE8686902772T priority patent/DE3685930T2/de
Priority claimed from AU56478/86A external-priority patent/AU592560B2/en
Publication of WO1986006487A1 publication Critical patent/WO1986006487A1/fr
Priority to DK624986A priority patent/DK165197C/da
Priority to NO865215A priority patent/NO168793C/no

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6878Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids in epitope analysis
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803General methods of protein analysis not limited to specific proteins or families of proteins

Definitions

  • This invention relates to a method of detecting or determining a sequence of monomer molecules which corresponds to the ligand molecule for a particular receptor.
  • the sequence of monomer molecules so determined is the mimotope (defined below) of the particular ligand.
  • the mimotope which is determined by this method may not have any obvious or direct relationship to the natural ligand molecule, but will share with it the ability ' to react with the receptor, and indeed, the mimotope so determined may be modified to incorporate specific or additional properties in the reaction with the receptor.
  • Such a mimotope could then be used to replace the natural ligand in the treatment or prevention of particular disease or it may be used to mediate a particular biological effect.
  • receptor a molecule or molecular complex which will combine specifically with its particular ligand molecule. It is those receptors which on binding with their particular ligand(s) mediate a biological function that are of most interest.
  • Examples of receptors include, but are not restricted to, the common class of receptors associated with the surface membrane of cells and include, for instance, the immunologically important receptors of B-cells, T-cells, macrophages and the like.
  • Another example is receptors for acetyl choline on nerve cells which cause a nerve pulse to be transmitted down the length of the neuron when the receptor molecule reacts with its ligand, acetyl choline.
  • epitope the specific surface of an antigen molecule which is delineated by the area of interaction with the sub-class of receptors known as antibodies.
  • catamer a polymer molecule which is a precisely defined linear sequence formed by the condensation of small molecules. Note that this term includes molecules in which different types of condensation reactions are used to join the small molecules. A number prefixed to the word "catamer” implies that the catamer is formed by the condensation of the indicated number of small molecules, for example, 8-catamer means that the catamer is made up from eight small molecules. Examples of catamers include any given peptide and any given oligo-saccharide.
  • the monomer a member of the set of small molecules which can be condensed together to form a catamer.
  • the set of monomers includes but is not restricted to, for example, the set of common L-amino acids, the set of D-amino acids, the set of synthetic amino acids, the set of nucleotides, and the set of pentoses and hexoses.
  • peptide a catamer in which the small molecules are alpha-amino acids and which are joined together through a peptide bond.
  • amino acids may be the L-optical.isomer or the D-optical isomer.
  • mimotope a catamer which in at least one of its conformations has a surface region with the equivalent molecular topology to the binding surface of the ligand molecule of which it is the mimic. In the context of immunological ⁇ receptors, the mimotope mimics the. epitope of the antigen.
  • complementary refers to the matching together of the reacting surfaces of an ligand molecule and its receptor.
  • the receptor and its ligand can be described as complementary, and furthermore, the contact surface characteristics are complementary to each other.
  • paratope the combining site of an antibody which is complementary to a particular epitope.
  • ligand molecule is the molecule which binds to a particular receptor and when bound to it mediates the biological function associated with that particular receptor.
  • receptors which can be investigated by this method include, but are not restricted to:
  • hormone receptors for instance the receptors for insulin and Growth Hormone; determination of the mimotopes of the ligands binding to these receptors may lead to the development of an oral replacement of the daily injections which diabetics must take to relieve the symptoms of diabetes, and in the other case, a replacement for the scarce human Growth Hormone which can only be obtained from cadavers or by recombinant DNA technology.
  • Other examples are the vasoconstrictive hormone receptors; determination of the mimotope of the ligand binding to these receptors may lead to the ' development of drugs to control blood pressure.
  • opiate receptors determination of mimotopes of the ligands binding to the opiate-receptors in the brain may lead to the development of less-addictive replacements for morphine and related drugs.
  • microorganism receptors determination of mimotopes of the ligands binding to a receptor such as specific transport proteins or enzymes essential to survival to microorganisms, may lead to a new class of antibiotics. Of particular value would be antibiotics against protozoa and those bacteria resistant to the antibiotics in current use.
  • enzymes for instance, the enzymes responsible for cleaving neural transmitters; determination of mimotopes able to modulate the action of the enzymes which cleave the different neural transmitters may lead to the development of drugs which can be used in the treatment of disorders of neural transmission; and,
  • antibodies for instance the ligand-binding site on the antibody molecule which combines with the epitope of an antigen of interest; determination of a. mimotope for the epitope may lead to the development of vaccines of which the immunogen is based on one or more mimotopes or diagnostic agents or compounds useful in the therapeutic treatment of autoimmune diseases.
  • ligands which can be investigated by this method include, but are not restricted to:
  • toxins and venoms for instance, the combining site of the toxin molecule which reacts with a particular receptor in the body to give the particular symptom(s) of intoxication; determination of mimotopes to the combining site of the ligand may lead to the development of drugs which can be used to treat envenomation by snakes and other poisonous animals without the side effects of heterologous antivenenes.
  • virus and other microorganism capsid molecules for instance, the combining site on the virus coat molecule which reacts with a particular receptor on the cell membrane in the body and which allows the virus to invade and thus infect the particular cell; determination of mimotopes to this combining site may lead to the development of drugs which specifically prevent intracellular invasion by the virus and thus prevent their replication.
  • the most usual group* of small molecules which may be condensed together to form a catamer is the group of alpha-amino acids.
  • other molecules which are consistent with a different chosen chemistry may also be used; for example, catamers formed from the specified sequential condensation of nucleotides or saccharides.
  • Another group of small molecules would be the non-genetically coded amino acids such as beta-amino acids, which may be used to advantage to add an additional bond at specified positions within the catamer.
  • the method of the present invention is based on the realisation that a given receptor will specifically react with a catamer which is the mimotope for the ligand to which the receptor is directed. It further relies on modern techniques of immunology to detect reaction between a receptor and its ligand when both are present.
  • reaction is readily detected between a receptor and short mimotopes. These short mimotopes when condensed together then bind to the receptor with either a greater affinity or with greater specificity for that receptor. This reaction can be detected even when the short mimotope presented to the receptor is as small as two monomer molecules long.
  • the final structure of a strongly-binding mimotope can be determined.
  • a method of detecting or determining the sequence of monomers which is a topographical equivalent of the ligand which is complementary to the particular receptor of interest and prior knowledge is irrelevant about the identity, structure and sequence of the receptor or its ligand. The method comprises the steps of:-
  • D 2- D l wherein D» represents a designated monomer selected from a first set of monomers, and D ⁇ represents a designated monomer selected from a second set of monomers which may be the same as or different to said first set of monomers; said plurality of catamers comprising catamers in which each designated monomer is systematically varied to contain members from the respective set of monomers;
  • the method may also comprise the further steps of:
  • D. and D 2 are as defined above, and preferably represent a combination of monomers corresponding to a catamer which binds to said receptor, and D, represents a designated monomer selected from a third set of monomers which may be the same as or different to either the first or the second set of monomers, and
  • steps a. and b. above may be repeated to further "extend" the catamers by systematically adding further monomers to the catamers, and testing in the same manner as in step b, above.
  • the metho ' d of this invention may comprise the steps of:
  • D 2 -Sp- Dl wherein D, and D ? are as defined above and Sp is a spacer molecule which can modify the relative orientation of the monomers D 1 and D 2 ;
  • the spacer molecule "Sp” as described above may also be systematically introduced into all possible positions of the "extended” catamers referred to above, and tested in the same manner as in Step B. above.
  • the method of the invention may further comprise the steps of «. systematically replacing the monomers in any of the catamers referred to above with their optical isomers, either individually or in combinations of monomers, and again performing steps 2 and 3 as describe above.
  • the method may further comprise the steps of systematically replacing the monomers in any of the catamers referred to above which bind with the receptor of interest, either individually or in combinations of monomers, with other monomers selected from the respective set(s) of monomers, and again performing steps 2 and 3 as described above.
  • the method of this invention requires no previous information about the nature of the ligand and in particular it requires no foreknowledge of the sequence of monomers which make up the ligand. In fact, it is not necessary for the application of this invention to know the source or identity of the ligand to which the receptor is directed. Furthermore, this invention makes no assumptions about the nature of the ligand of the particular receptor. This method will identify mimotopes of discontinuous as well as continuous ligands. Because of the very nature of the method of the invention it will be appreciated that mimotopes may or may not consist of members of the same set of monomers which make up the ligand of which it is the mimic.
  • the plurality of catamers required to be synthesized in the application of this invention may be prepared by any of the known method of catamer synthesis.
  • the preferred methods when the monomers are amino acids is to use the solid phase technique described in Australian Patent Specification No. 25429/84, whereby each catamer is synthesized on a polyethylene support.
  • the method of the present invention is carried out by screening a plurality of synthesized catamers against the antibody of interest.
  • the antibody will be a monoclonal antibody which can be prepared by any of the usual methods. Polyclonal antiserum from humans and animals may be used if a source of monoclonal antibody with the desired characteristics is not available, however, analysis of the resulting data may be complicated because the reaction observed could be from more than one monoclonal antibody.
  • polyclonal antiserum it may be necessary to reduce the antibody diversity by using techniques well known to those skilled in the art, for example iso-electric focusing, HPLC-based chromatography or affinity chromatography.
  • Lk represents a linker molecule which provides a suitable end group for condensing the monomers to the solid support.
  • D and “D.-” represent designated 0 positions occupied by monomers which are selected from known sets of monomers; but which are altered systematically between catamers. It should be noted that the set of monomers used for the D. designated position need not be the same set of monomers used for c the D 2 designated position.
  • Y in the general formula is an end group of the catamer and may be, but is not restricted to, for example a hydrogen atom or an acetyl group. "Y” may also be another molecule which is coupled to the catamer to preserve particular Q characteristics of the molecular environment of a peptide bond at the amino terminal designated position.
  • j . is the number of members in the set of monomers to be coupled in the D.. position and jj is the 5 number of members in the set of monomers to be coupled in the D ⁇ position then a total of ⁇ . j_ different catamers will be synthesized.
  • the support rods are prepared so that the monomers can be coupled to them by coupling an appropriate linker molecule.
  • each rod 0 will be treated with a reaction mixture which contains only a single monomer such as a protected amino acid or the like.
  • each of the i. monomers are coupled to j. rods.
  • each rod is treated with a reaction mixture which 5 contains a single monomer such as a protected amino acid or the like.
  • Each of the j. rods which has a particular monomer in the D.. position will have a different monomer coupled at the D position. In this way every combination of the members of the set(s) of monomers Q - will be found in the i.. . rods.
  • the desired end group, "Y” is then coupled using the appropriate chemistry.
  • Sp is a spacer molecule which may restrict the relative orientation of the monomers at the designated positions to a particular geometrical configuration(s) .
  • the spacer molecule may also be deliberately chosen to allow a greater flexibility to the relative geometric configuration between monomers in the designated positions, D.. and D . It should be noted that members of the set of spacer molecules may be made up from the condensation of more than one monomer.
  • spacer molecules include, but are not restricted to g ⁇ ycine (approximately linear extension) , beta-alanine (increased flexibility) , proline (forced bend) , glycyl-proline (extended bend, otherwise known as reverse bend in protein structure terminology) and o-aminobenzoic acid (a planar bend) .
  • the plurality of catamers prepared as in A. above are then contacted with the particular antibody of interest.
  • the reaction between antibody and each catamer can then be detected by any of the usual methods, for example, radioimmunoassay (RIA) .
  • RIA radioimmunoassay
  • the preferred method of detection is to use the well known enzyme-linked im unosorbent assay (ELISA) .
  • each assay antibodies can be removed from the catamers by, for example, washing with a solution of 8M urea, 0.1% 2-mercaptoethanol and 0.1% sodium dodecylsulphate followed by several washes in phosphate buffered saline. In this way the plurality of catamers may be used for testing with many other antibodies.
  • the selected catamers can be synthesized using similar methods to those used in A. After the selected catamers have been synthesized they are reacted with the 10 antibody of interest. It is then simple to select the catamer which binds most strongly with the antibody.
  • the binding of the mimotope with the antibody can be further enhanced by synthesizing a further
  • This plurality of catamers consists of adding spacer molecules (-Sp-) systematically at all positions; of the mimotope; and where feasible systematically replacing each monomer of
  • the mimotopes are being determined for the antigen against which different monoclonal antibodies were raised.
  • the defined set of monomers is the set of the common L-alpha
  • the linker molecule, -Lk- was 3 amino-N (6-aminohexyl)-propanamide and the end group, Y-, was the acetyl moiety.
  • the mimotope is determined for an antigenic determinant of human chorionic gonadotrophin which
  • the linker molecule, -Lk- is the same as that used in earlier Examples.
  • the end group, Y- is either ⁇ -alanyl-3-alanine or ⁇ -alanine.
  • the defined set of monomers was extended to include the D-optical isomers of the common alpha amino acids and several unusual amino acids. In Example 6 below, the mimotope is determined for a receptor site on viruses.
  • the linker molecule, -Lk- is the same as for earlier Examples.
  • the end group, Y- is either 0-alanyl- ⁇ -alanine or 8-alanine.
  • the defined set of monomers was extended as described for Example 5.
  • a monoclonal antibody was raised against sperm whale myoglobin using the usual techniques. This monoclonal antibody was tested against a set of catamers with the general formula:
  • a further set of catamers was synthesized which comprised all 4-catamers which could be made from the set of monomers; Glutamic acid (E) , Phenylalanine (F) , Histidine (H) and Leucine (L) .
  • the following catamers were found to bind significantly higher than the remaining catamers:-
  • a monoclonal ⁇ ntibody was raised against sperm whale myoglobin using the usual techniques. This monoclonal antibody was tested against a set of catamers with the general formula:
  • the structure below is a two dimensional representation of the spatial relationship between the amino acids, F, E, L, and H when bound to a monoclonal antibody against sperm whale myoglobin (see Example 2) . It must be noted * that this is an illustration only and must not be interpreted to mean that the catamer illustrated is planar. Furthermore, the bonds joining F to E and L to H are meant to represent a distance greater than that of a peptide bond.
  • a mimotope to a monoclonal antiserum raised against Foot and Mouth Disease Virus was delineated to the sequence W-Q-M-G-H-S.
  • a series of catamers were synthesized in which a beta-alanine residue was introduced systematically between monomers.
  • the sequence W-Q-M- ⁇ -G-H-S gave a response which was significantly larger than the base sequence, where ⁇ represents beta-alanine. It was further found that excellent binding to the antibody was achieved with the sequences:
  • the mimotope is made up of two parts; furthermore, the joining together of these parts is not critical for the mimotope to be able to react strongly with the antibody.
  • a monoclonal antibody (S218-4) was raised against human chorionic gonadotrophin using the usual techniques. This was tested with catamers with the general formula:-
  • the set of monomers used in the designated position D was extended to 'include both the L- and D- optical isomers of the common alpha amino acids, and the unusual amino acids, ⁇ -amino butyric acid, ⁇ -amino butyric acid, L-norcleucine, sarcosine, ornithine, L-norvaline, L-homophenylalanine, ⁇ -alanine.
  • these sets of catamers were reacted with the monoclonal antibody it was found that the catamers which reacted strongly were:-
  • This Example illustrates the application of the invention to the detection of a mimotope to a receptor which is not an antigen to which an antibody has been raised.
  • This Example detects mimotopes of a receptor to which a virion binds.
  • the test system for detecting binding to catamers has to be modified.
  • the synthesized catamers are allowed to react with influenza virus particles (strain A-Shearwater/1/72) . After reaction the catamers are washed to remove unbound virus particles. The presence of bound virus particles was detected by reacting with a polyclonal antibody (S227-1) which had been raised against the hae aglutinin of influenza virus A/Shearwater/1/72. Antibodies which had reacted with the bound virus were detected in the usual way by ELISA.
  • a set of catamers was synthesized with the general formula:- Y- j -D--D-. -Lk-(solid support] where the end group, Y.. -, is ⁇ -alanyl- ⁇ -alanine.
  • the set of monomers which was used in the designated positions D. and D fatigue consisted of the D- and L- optical isomers of the common alpha amino acids.
  • the catamers After removal of the bound virus from the catamers, the catamers were reacted with the polyclonal antibody S227-1 at the same concentration as that used to detect bound virus to ensure that the peaks found were due to binding of virus rather than binding of the polyclonal antibody.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Pathology (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Biophysics (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Méthode permettant de détecter ou de déterminer la séquence de monomères qui est un équivalent topographique du liant qui est complémentaire à un récepteur particulier présentant un intérêt, ladite méthode comprenant les étapes suivantes: 1. synthétisation d'une pluralité de catamères, chacun desdits catamères faisant partie de la formule générale D2-D1, D1 représentant un monomère désigné sélectionné dans un premier groupe de monomères, et D2 représentant un monomère désigné sélectionné dans un second groupe de monomères qui peut être égal au premier groupe de monomères ou différent; ladite pluralité de catamères comprenant des catamères dans lesquels chaque monomère désigné varie systématiquement de manière à comprendre des éléments du groupe respectif de monomères; 2. mise en contact de chaque catamère avec le récepteur présentant un intêret; et 3. détection ou déterminatin de la présence ou de l'absence de liants entre chaque catamère et ledit récepteur. La méthode peut également se rapporter à la synthèse d'autres pluralités de catamères, comprenant des catamères contenant des molécules intermédiaires, de manière à former une séquence de monomères qui est le mimotope du liant.
PCT/AU1986/000110 1985-04-22 1986-04-22 Methode de determination de mimotopes WO1986006487A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
AT86902772T ATE78097T1 (de) 1985-04-22 1986-04-22 Verfahren zur bestimmung von mimitopen.
DE8686902772T DE3685930T2 (de) 1985-04-22 1986-04-22 Verfahren zur bestimmung von mimitopen.
DK624986A DK165197C (da) 1985-04-22 1986-12-22 Fremgangsmaade til bestemmelse af mimotoper
NO865215A NO168793C (no) 1985-04-22 1986-12-22 Fremgangsmaate for paavisning eller bestemmelse av mimotoper

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
AUPH024085 1985-04-22
AUPH0240/85 1985-04-22
AU56478/86A AU592560B2 (en) 1985-04-22 1986-04-22 Improved method for determining mimotopes

Publications (1)

Publication Number Publication Date
WO1986006487A1 true WO1986006487A1 (fr) 1986-11-06

Family

ID=25631326

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/AU1986/000110 WO1986006487A1 (fr) 1985-04-22 1986-04-22 Methode de determination de mimotopes

Country Status (1)

Country Link
WO (1) WO1986006487A1 (fr)

Cited By (53)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0314402A2 (fr) 1987-10-26 1989-05-03 Schering Biotech Corporation Anticorps monoclonaux contre l'interleukine-4 humaine et hybridomes qui les produisent
FR2631451A1 (fr) * 1988-05-13 1989-11-17 Inst Nat Sante Rech Med Procede de caracterisation de l'epitope intervenant dans une reaction antigene anticorps, kit ou necessaire pour la mise en oeuvre du procede
EP0390612A1 (fr) 1989-03-31 1990-10-03 Synbiotics Corporation Production de médicament utilisant des anticorps à réactions non croisées
WO1991006356A1 (fr) * 1989-10-31 1991-05-16 Terrapin Technologies, Inc. Procede d'identification de ligands de liaison d'analytes
US5143854A (en) * 1989-06-07 1992-09-01 Affymax Technologies N.V. Large scale photolithographic solid phase synthesis of polypeptides and receptor binding screening thereof
US5182366A (en) * 1990-05-15 1993-01-26 Huebner Verena D Controlled synthesis of peptide mixtures using mixed resins
WO1994002225A1 (fr) * 1992-07-27 1994-02-03 Terrapin Technologies, Inc. Familles de sorbants
US5300425A (en) * 1987-10-13 1994-04-05 Terrapin Technologies, Inc. Method to produce immunodiagnostic reagents
US5340474A (en) * 1988-03-24 1994-08-23 Terrapin Technologies, Inc. Panels of analyte-binding ligands
US5409611A (en) * 1988-03-24 1995-04-25 Terrapin Technoogies, Inc. Method to identify analyte-binding ligands
US5422426A (en) * 1991-06-18 1995-06-06 Eli Lilly And Company Rapid synthesis and screening of peptide mimetics
US5438119A (en) * 1988-05-02 1995-08-01 The Regents Of The University Of California Method of obtaining a peptide with desired target property
US5489678A (en) * 1989-06-07 1996-02-06 Affymax Technologies N.V. Photolabile nucleoside and peptide protecting groups
US5492807A (en) * 1993-11-19 1996-02-20 Santi; Daniel V. Method of obtaining diagnostic reagents, assays and therapeutics based on clinical manifestations of a disease
US5498538A (en) * 1990-02-15 1996-03-12 The University Of North Carolina At Chapel Hill Totally synthetic affinity reagents
US5510240A (en) * 1990-07-02 1996-04-23 The Arizona Board Of Regents Method of screening a peptide library
EP0710724A2 (fr) 1994-10-06 1996-05-08 Akzo Nobel N.V. Antigènes de Toxoplasma gondii
WO1996024610A1 (fr) * 1995-02-06 1996-08-15 Chiron Mimotopes Pty. Ltd. Banques combinatoires de tripeptides ramifies utiles pour traiter les troubles lies a l'activateur du plasminogene urokinase
EP0773227A1 (fr) * 1991-09-18 1997-05-14 Affymax Technologies N.V. Collections d'oligomères diverses, utiles pour préparer des médicaments, réactifs diagnostiques pesticides et herbicides
EP0775746A2 (fr) 1995-07-03 1997-05-28 Akzo Nobel N.V. Vaccin contre la coccidiose aviaire
US5700637A (en) * 1988-05-03 1997-12-23 Isis Innovation Limited Apparatus and method for analyzing polynucleotide sequences and method of generating oligonucleotide arrays
US5747334A (en) * 1990-02-15 1998-05-05 The University Of North Carolina At Chapel Hill Random peptide library
US5763570A (en) * 1987-10-13 1998-06-09 Terrapin Technologies, Inc. Glutathione analogs and paralog panels comprising glutathione mimics
US5840841A (en) * 1990-05-15 1998-11-24 Chiron Corporation Method and apparatus for biopolymer synthesis
US5866363A (en) * 1985-08-28 1999-02-02 Pieczenik; George Method and means for sorting and identifying biological information
US5908919A (en) * 1993-10-01 1999-06-01 Terrapin Technologies Urethane mediated, GST specific molecular release systems
US5955432A (en) * 1992-04-03 1999-09-21 Terrapin Technologies, Inc. Metabolic effects of certain glutathione analogs
US5958792A (en) * 1995-06-07 1999-09-28 Chiron Corporation Combinatorial libraries of substrate-bound cyclic organic compounds
US5994083A (en) * 1993-05-11 1999-11-30 Istituto Di Recerche Di Biologia Molecolare P. Angeletti S.P.A. Process for the preparation of immunogens or diagnostic reagents, and immunogens or diagnostic reagents thereby obtainable
US6054270A (en) * 1988-05-03 2000-04-25 Oxford Gene Technology Limited Analying polynucleotide sequences
US6054438A (en) * 1987-01-07 2000-04-25 Imperial Cancer Research Technology Limited Nucleic acid fragments encoding portions of the core protein of the human mammary epithelial mucin
US6075121A (en) * 1990-05-15 2000-06-13 Chiron Corporation Modified peptide and peptide libraries with protease resistance, derivatives thereof and methods of producing and screening such
US6660276B1 (en) 1994-02-16 2003-12-09 The University Of Virginia Patent Foundation Peptides recognized by melanoma-specific cytotoxic lymphocytes, and uses therefor
US6849462B1 (en) 1991-11-22 2005-02-01 Affymetrix, Inc. Combinatorial strategies for polymer synthesis
US6919211B1 (en) 1989-06-07 2005-07-19 Affymetrix, Inc. Polypeptide arrays
US6955915B2 (en) 1989-06-07 2005-10-18 Affymetrix, Inc. Apparatus comprising polymers
US7056666B2 (en) 1990-12-06 2006-06-06 Affymetrix, Inc. Analysis of surface immobilized polymers utilizing microfluorescence detection
US7378236B1 (en) 1994-06-17 2008-05-27 The Board Of Trustees Of The Leland Stanford Junior University Method for analyzing gene expression patterns
US7442499B2 (en) 1994-06-17 2008-10-28 The Board Of Trustees Of The Leland Stanford Junior University Substrates comprising polynucleotide microarrays
US7625697B2 (en) 1994-06-17 2009-12-01 The Board Of Trustees Of The Leland Stanford Junior University Methods for constructing subarrays and subarrays made thereby
US7691330B1 (en) 1991-11-22 2010-04-06 Affymetrix, Inc. Combinatorial strategies for polymer synthesis
US7811751B2 (en) 1988-05-03 2010-10-12 Oxford Gene Technology Limited Analysing polynucleotide sequences
US7888494B2 (en) 1988-05-03 2011-02-15 Oxford Gene Therapy Limited Analysing polynucleotide sequences
WO2012089748A1 (fr) 2010-12-29 2012-07-05 Intervet International B.V. Antigène vaccinal contre la babésiose canine
WO2012143488A1 (fr) 2011-04-21 2012-10-26 The University Court Of The University Of Aberdeen Protéine destinée à être utilisée comme médicament
WO2013034682A1 (fr) 2011-09-08 2013-03-14 Umc Utrecht Holding B.V. Vaccin basé sur la protéine analogue à un superantigène staphylococcique 3 (ssl3)
WO2013113865A1 (fr) 2012-02-03 2013-08-08 The Pirbright Institute Vaccin à vecteur eimeria contre le campylobacter jejuni
WO2014114812A1 (fr) 2013-01-28 2014-07-31 Danmarks Tekniske Universitet Vaccin à sous-unité contre mycobacterium avium subsp. paratuberculosis à une ou plusieurs étapes
US9366667B2 (en) 2009-10-02 2016-06-14 The Trustees Of Columbia University In The City Of New York Piscine reovirus diagnostic compositions
EP3085367A2 (fr) 2007-03-20 2016-10-26 Brandeis University Compositions pour le diagnostic, le traitement et la prévention de la sclérose latérale amyotrophique et apparentées
WO2021099444A1 (fr) 2019-11-20 2021-05-27 Intervet International B.V. Nouveau vaccin contre heamophilus parasuis
WO2021099446A1 (fr) 2019-11-20 2021-05-27 Intervet International B.V. Nouveau vaccin contre heamophilus parasuis
WO2021099458A1 (fr) 2019-11-20 2021-05-27 Intervet International B.V. Nouveau vaccin contre haemophilus parasuis

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1984002983A1 (fr) * 1983-01-21 1984-08-02 Frederick James Primus Antigenes specifiques de la famille cea (antigene carcino-embryonaire), anticorps specifiques de ces antigenes et leurs procedes d'utilisation
WO1984003564A1 (fr) 1983-03-08 1984-09-13 Commw Serum Lab Commission Procede de determination de sequences d'acides amines antigeniquement actives
WO1984003506A1 (fr) 1983-03-08 1984-09-13 Commw Serum Lab Commission Sequences d'acides amines antigeniquement actives
AU2542884A (en) * 1983-03-08 1984-09-20 Chiron Mimotopes Pty Ltd Immunogenic deteminants of foot and mouth disease virus
WO1985002121A1 (fr) * 1983-11-07 1985-05-23 The Wistar Institute Reponse immune aux tumeurs et virus induits par des anticorps anti-idiotypes
AU3656084A (en) * 1983-12-12 1985-06-20 Miles Laboratories Inc. Nucleic acid hybridization assay employing antibodies to intercalation complexes
AU3656384A (en) * 1983-12-12 1985-06-20 Miles Laboratories Inc. Hybridization assay employing labeledprobe and anti-hybrid
AU3656284A (en) * 1983-12-12 1985-06-20 Miles Laboratories Inc. Hybridization assay with immobilization of hybrids by anti- hybrid binding
AU4533985A (en) 1984-07-24 1986-01-30 Coselco Mimotopes Pty Ltd Method for determining mimotopes

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1984002983A1 (fr) * 1983-01-21 1984-08-02 Frederick James Primus Antigenes specifiques de la famille cea (antigene carcino-embryonaire), anticorps specifiques de ces antigenes et leurs procedes d'utilisation
WO1984003564A1 (fr) 1983-03-08 1984-09-13 Commw Serum Lab Commission Procede de determination de sequences d'acides amines antigeniquement actives
WO1984003506A1 (fr) 1983-03-08 1984-09-13 Commw Serum Lab Commission Sequences d'acides amines antigeniquement actives
AU2542884A (en) * 1983-03-08 1984-09-20 Chiron Mimotopes Pty Ltd Immunogenic deteminants of foot and mouth disease virus
WO1985002121A1 (fr) * 1983-11-07 1985-05-23 The Wistar Institute Reponse immune aux tumeurs et virus induits par des anticorps anti-idiotypes
AU3656084A (en) * 1983-12-12 1985-06-20 Miles Laboratories Inc. Nucleic acid hybridization assay employing antibodies to intercalation complexes
AU3656384A (en) * 1983-12-12 1985-06-20 Miles Laboratories Inc. Hybridization assay employing labeledprobe and anti-hybrid
AU3656284A (en) * 1983-12-12 1985-06-20 Miles Laboratories Inc. Hybridization assay with immobilization of hybrids by anti- hybrid binding
AU4533985A (en) 1984-07-24 1986-01-30 Coselco Mimotopes Pty Ltd Method for determining mimotopes

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
EMBO J., vol. 3, no. 6, 1984, pages 1295 - 1300
PNAS, vol. 82, 1985, pages 178 - 182
PNAS, vol. 87, 1984, pages 3998 - 4002
Scientific American, February 1983 pages 48-56 (page 50 lines 33-39), R.A. LERNER, "Synthetic Vaccines" *
See also references of EP0220245A4 *

Cited By (81)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6605448B1 (en) 1985-08-28 2003-08-12 George Pieczenik Method and means for sorting and identifying biological information
US5866363A (en) * 1985-08-28 1999-02-02 Pieczenik; George Method and means for sorting and identifying biological information
US6054438A (en) * 1987-01-07 2000-04-25 Imperial Cancer Research Technology Limited Nucleic acid fragments encoding portions of the core protein of the human mammary epithelial mucin
US5763570A (en) * 1987-10-13 1998-06-09 Terrapin Technologies, Inc. Glutathione analogs and paralog panels comprising glutathione mimics
US5541070A (en) * 1987-10-13 1996-07-30 Kauvar; Lawrence M. Method to identify and characterize candidate drugs
US5300425A (en) * 1987-10-13 1994-04-05 Terrapin Technologies, Inc. Method to produce immunodiagnostic reagents
EP0314402A2 (fr) 1987-10-26 1989-05-03 Schering Biotech Corporation Anticorps monoclonaux contre l'interleukine-4 humaine et hybridomes qui les produisent
US5133866A (en) * 1988-03-24 1992-07-28 Terrapin Technologies, Inc. Method to identify analyte-bending ligands
US5340474A (en) * 1988-03-24 1994-08-23 Terrapin Technologies, Inc. Panels of analyte-binding ligands
US5409611A (en) * 1988-03-24 1995-04-25 Terrapin Technoogies, Inc. Method to identify analyte-binding ligands
US5567317A (en) * 1988-03-24 1996-10-22 Terrapin Technologies, Inc. Method to identify analyte-binding ligands
US5438119A (en) * 1988-05-02 1995-08-01 The Regents Of The University Of California Method of obtaining a peptide with desired target property
US5641862A (en) * 1988-05-02 1997-06-24 The Regents Of The University Of California General method for producing and selecting peptides with specific properties
US7811751B2 (en) 1988-05-03 2010-10-12 Oxford Gene Technology Limited Analysing polynucleotide sequences
US6054270A (en) * 1988-05-03 2000-04-25 Oxford Gene Technology Limited Analying polynucleotide sequences
US7888494B2 (en) 1988-05-03 2011-02-15 Oxford Gene Therapy Limited Analysing polynucleotide sequences
US5700637A (en) * 1988-05-03 1997-12-23 Isis Innovation Limited Apparatus and method for analyzing polynucleotide sequences and method of generating oligonucleotide arrays
FR2631451A1 (fr) * 1988-05-13 1989-11-17 Inst Nat Sante Rech Med Procede de caracterisation de l'epitope intervenant dans une reaction antigene anticorps, kit ou necessaire pour la mise en oeuvre du procede
EP0390612A1 (fr) 1989-03-31 1990-10-03 Synbiotics Corporation Production de médicament utilisant des anticorps à réactions non croisées
US5889165A (en) * 1989-06-07 1999-03-30 Affymetrix, Inc. Photolabile nucleoside protecting groups
US5744305A (en) * 1989-06-07 1998-04-28 Affymetrix, Inc. Arrays of materials attached to a substrate
US5143854A (en) * 1989-06-07 1992-09-01 Affymax Technologies N.V. Large scale photolithographic solid phase synthesis of polypeptides and receptor binding screening thereof
US6955915B2 (en) 1989-06-07 2005-10-18 Affymetrix, Inc. Apparatus comprising polymers
US7087732B2 (en) 1989-06-07 2006-08-08 Affymetrix, Inc. Nucleotides and analogs having photoremovable protecting groups
US6124102A (en) * 1989-06-07 2000-09-26 Affymetrix, Inc. Methods for determining receptor-ligand binding using probe arrays
US5744101A (en) * 1989-06-07 1998-04-28 Affymax Technologies N.V. Photolabile nucleoside protecting groups
US6600031B1 (en) * 1989-06-07 2003-07-29 Affymetrix, Inc. Methods of making nucleic acid or oligonucleotide arrays
US6919211B1 (en) 1989-06-07 2005-07-19 Affymetrix, Inc. Polypeptide arrays
US5489678A (en) * 1989-06-07 1996-02-06 Affymax Technologies N.V. Photolabile nucleoside and peptide protecting groups
WO1991006356A1 (fr) * 1989-10-31 1991-05-16 Terrapin Technologies, Inc. Procede d'identification de ligands de liaison d'analytes
AU654940B2 (en) * 1989-10-31 1994-12-01 Telik, Inc. Method to identify analyte-binding ligands
US5498538A (en) * 1990-02-15 1996-03-12 The University Of North Carolina At Chapel Hill Totally synthetic affinity reagents
US5747334A (en) * 1990-02-15 1998-05-05 The University Of North Carolina At Chapel Hill Random peptide library
US5948635A (en) * 1990-02-15 1999-09-07 University Of North Carolina At Chapel Hill Totally Synthetic Affinity Reagents
US5844076A (en) * 1990-02-15 1998-12-01 The University Of North Carolina At Chapel Hill Totally synthetic affinity reagents
US5852167A (en) * 1990-02-15 1998-12-22 The University Of North Carolina At Chapel Hill Totally synthetic affinity reagents
US5625033A (en) * 1990-02-15 1997-04-29 The University Of North Carolina At Chapel Hill Totally synthetic affinity reagents
US6075121A (en) * 1990-05-15 2000-06-13 Chiron Corporation Modified peptide and peptide libraries with protease resistance, derivatives thereof and methods of producing and screening such
US5182366A (en) * 1990-05-15 1993-01-26 Huebner Verena D Controlled synthesis of peptide mixtures using mixed resins
US5840841A (en) * 1990-05-15 1998-11-24 Chiron Corporation Method and apparatus for biopolymer synthesis
US5650489A (en) * 1990-07-02 1997-07-22 The Arizona Board Of Regents Random bio-oligomer library, a method of synthesis thereof, and a method of use thereof
US5858670A (en) * 1990-07-02 1999-01-12 The Arizona Board Of Regents Bio-oligomer libraries and a method of use thereof
US5510240A (en) * 1990-07-02 1996-04-23 The Arizona Board Of Regents Method of screening a peptide library
US7056666B2 (en) 1990-12-06 2006-06-06 Affymetrix, Inc. Analysis of surface immobilized polymers utilizing microfluorescence detection
US7329496B2 (en) 1990-12-06 2008-02-12 Affymetrix, Inc. Sequencing of surface immobilized polymers utilizing microflourescence detection
US5422426A (en) * 1991-06-18 1995-06-06 Eli Lilly And Company Rapid synthesis and screening of peptide mimetics
EP0773227A1 (fr) * 1991-09-18 1997-05-14 Affymax Technologies N.V. Collections d'oligomères diverses, utiles pour préparer des médicaments, réactifs diagnostiques pesticides et herbicides
US6864101B1 (en) 1991-11-22 2005-03-08 Affymetrix, Inc. Combinatorial strategies for polymer synthesis
US6849462B1 (en) 1991-11-22 2005-02-01 Affymetrix, Inc. Combinatorial strategies for polymer synthesis
US7691330B1 (en) 1991-11-22 2010-04-06 Affymetrix, Inc. Combinatorial strategies for polymer synthesis
US7736906B2 (en) 1991-11-22 2010-06-15 Affymetrix, Inc. Combinatorial strategies for polymer synthesis
US5955432A (en) * 1992-04-03 1999-09-21 Terrapin Technologies, Inc. Metabolic effects of certain glutathione analogs
US6013462A (en) * 1992-04-03 2000-01-11 Terrapin Technologies, Inc. Glutathione analogs as reagents
US5801225A (en) * 1992-07-27 1998-09-01 Terrapin Technologies, Inc. Sorbent families
WO1994002225A1 (fr) * 1992-07-27 1994-02-03 Terrapin Technologies, Inc. Familles de sorbants
US5599901A (en) * 1992-07-27 1997-02-04 Terrapin Technologies, Inc. Sorbent families
US5994083A (en) * 1993-05-11 1999-11-30 Istituto Di Recerche Di Biologia Molecolare P. Angeletti S.P.A. Process for the preparation of immunogens or diagnostic reagents, and immunogens or diagnostic reagents thereby obtainable
US6541210B1 (en) 1993-05-11 2003-04-01 Istituto Di Recerche Di Biologia Moleculare P. Angeletti S.P.A. Process for the preparation of immunogens or diagnostic reagents, and immunogens or diagnostic reagents thereby obtainable
US5908919A (en) * 1993-10-01 1999-06-01 Terrapin Technologies Urethane mediated, GST specific molecular release systems
US5670312A (en) * 1993-11-19 1997-09-23 Santi; Daniel V. Method of obtaining diagnostic reagents, assays and therapeutics based on clinical manifestations of a disease
US5492807A (en) * 1993-11-19 1996-02-20 Santi; Daniel V. Method of obtaining diagnostic reagents, assays and therapeutics based on clinical manifestations of a disease
US6660276B1 (en) 1994-02-16 2003-12-09 The University Of Virginia Patent Foundation Peptides recognized by melanoma-specific cytotoxic lymphocytes, and uses therefor
US7378236B1 (en) 1994-06-17 2008-05-27 The Board Of Trustees Of The Leland Stanford Junior University Method for analyzing gene expression patterns
US7442499B2 (en) 1994-06-17 2008-10-28 The Board Of Trustees Of The Leland Stanford Junior University Substrates comprising polynucleotide microarrays
US7625697B2 (en) 1994-06-17 2009-12-01 The Board Of Trustees Of The Leland Stanford Junior University Methods for constructing subarrays and subarrays made thereby
EP0710724A2 (fr) 1994-10-06 1996-05-08 Akzo Nobel N.V. Antigènes de Toxoplasma gondii
WO1996024610A1 (fr) * 1995-02-06 1996-08-15 Chiron Mimotopes Pty. Ltd. Banques combinatoires de tripeptides ramifies utiles pour traiter les troubles lies a l'activateur du plasminogene urokinase
US5627210A (en) * 1995-02-06 1997-05-06 Chiron Corporation Branched combinatorial libraries
US5958792A (en) * 1995-06-07 1999-09-28 Chiron Corporation Combinatorial libraries of substrate-bound cyclic organic compounds
EP0775746A2 (fr) 1995-07-03 1997-05-28 Akzo Nobel N.V. Vaccin contre la coccidiose aviaire
EP3085367A2 (fr) 2007-03-20 2016-10-26 Brandeis University Compositions pour le diagnostic, le traitement et la prévention de la sclérose latérale amyotrophique et apparentées
US9366667B2 (en) 2009-10-02 2016-06-14 The Trustees Of Columbia University In The City Of New York Piscine reovirus diagnostic compositions
US9395356B2 (en) 2009-10-02 2016-07-19 The National Veterinary Institute Piscine reovirus immunogenic compositions
WO2012089748A1 (fr) 2010-12-29 2012-07-05 Intervet International B.V. Antigène vaccinal contre la babésiose canine
WO2012143488A1 (fr) 2011-04-21 2012-10-26 The University Court Of The University Of Aberdeen Protéine destinée à être utilisée comme médicament
WO2013034682A1 (fr) 2011-09-08 2013-03-14 Umc Utrecht Holding B.V. Vaccin basé sur la protéine analogue à un superantigène staphylococcique 3 (ssl3)
WO2013113865A1 (fr) 2012-02-03 2013-08-08 The Pirbright Institute Vaccin à vecteur eimeria contre le campylobacter jejuni
WO2014114812A1 (fr) 2013-01-28 2014-07-31 Danmarks Tekniske Universitet Vaccin à sous-unité contre mycobacterium avium subsp. paratuberculosis à une ou plusieurs étapes
WO2021099444A1 (fr) 2019-11-20 2021-05-27 Intervet International B.V. Nouveau vaccin contre heamophilus parasuis
WO2021099446A1 (fr) 2019-11-20 2021-05-27 Intervet International B.V. Nouveau vaccin contre heamophilus parasuis
WO2021099458A1 (fr) 2019-11-20 2021-05-27 Intervet International B.V. Nouveau vaccin contre haemophilus parasuis

Similar Documents

Publication Publication Date Title
US4833092A (en) Method for determining mimotopes
WO1986006487A1 (fr) Methode de determination de mimotopes
US5194392A (en) Method of determining mimotopes
US5541070A (en) Method to identify and characterize candidate drugs
EP0818467B1 (fr) Réseau peptidique aligné et procédé rapide et rationnel de dépistage d'une liaison ou d'un site d'interaction d'une protéine en utilisant celui-ci
Dambinova et al. The presence of autoantibodies to N-terminus domain of GluR1 subunit of AMPA receptor in the blood serum of patients with epilepsy
JP5611831B2 (ja) リウマチ性関節炎自己抗体との免疫反応性合成ペプチド
Sioud et al. Selection of ligands for polyclonal antibodies from random peptide libraries: potential identification of (auto) antigens that may trigger B and T cell responses in autoimmune diseases
US5955582A (en) Antibody against a 3-aminophenylboronic-glycated protein complex and its use in an immunoassay
Atanassov et al. New Zealand white rabbits immunized with RNA‐complexed total histones develop an autoimmune‐like response
Fiordalisi et al. Site-Directed Mutagenesis of. kappa.-Bungarotoxin: Implications for Neuronal Receptor Specificity
IE881289L (en) A method for the selective immunological determination of¹intact procollagen peptide (Type III) and procollagen (Type¹III) in body fluids, and means for carrying it out
Van Regenmortel Structure of antigens
AU592560B2 (en) Improved method for determining mimotopes
EP0351410A1 (fr) Peptide de liaison de complement
Ruben et al. Antibodies to calmodulin during experimental Trypanosoma brucei rhodesiense infections in rabbits
Herfurth et al. Determination of peptide regions exposed at the surface of the bacterial ribosome with antibodies against synthetic peptides
Zegers et al. Immunological recognition of peptides in medicine and biology
JP4387945B2 (ja) 抗イディオタイプ抗体、該抗イディオタイプ抗体の作成方法、及び該抗イディオタイプ抗体を用いたイディオタイプ抗体の調製方法
JPH06293798A (ja) ヒトパルボウイルスb19のエピトープ関連ペプチド
US6800462B2 (en) Production of recombinant proteins in vivo and use for generating antibodies
WO1996006357A1 (fr) Criblage pour des composes a potentiel therapeutique
DE19534988A1 (de) Verfahren zur Herstellung und Anwendung synthetischer, biotinylierter Peptide
JPH05339295A (ja) 抗体の作成方法
JPH05308992A (ja) モノクローナル抗体

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): DK JP NO US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE FR GB IT LU NL SE

WWE Wipo information: entry into national phase

Ref document number: 1986902772

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1986902772

Country of ref document: EP

WWG Wipo information: grant in national office

Ref document number: 1986902772

Country of ref document: EP

点击 这是indexloc提供的php浏览器服务,不要输入任何密码和下载