EP1397687A2 - Procede de determination de peptides/proteines, spectrometrie de masse pour metaux et determination de la conductivite - Google Patents

Procede de determination de peptides/proteines, spectrometrie de masse pour metaux et determination de la conductivite

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Publication number
EP1397687A2
EP1397687A2 EP02750770A EP02750770A EP1397687A2 EP 1397687 A2 EP1397687 A2 EP 1397687A2 EP 02750770 A EP02750770 A EP 02750770A EP 02750770 A EP02750770 A EP 02750770A EP 1397687 A2 EP1397687 A2 EP 1397687A2
Authority
EP
European Patent Office
Prior art keywords
proteins
sample
peptides
determination
metals
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP02750770A
Other languages
German (de)
English (en)
Inventor
Reinhold Kiehl
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
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
Priority claimed from DE2001126697 external-priority patent/DE10126697A1/de
Application filed by Individual filed Critical Individual
Publication of EP1397687A2 publication Critical patent/EP1397687A2/fr
Withdrawn legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G01N33/6866—Interferon

Definitions

  • the invention is generally concerned with the determination of peptides / proteins.
  • the invention also relates to mass spectrometry for metals and conductivity detectors.
  • mass spectrometry for metals and conductivity detectors are disclosed in the present documents and within the scope of the invention, these are additionally independent inventions with, under certain circumstances, independent protection needs and protection options.
  • the invention relates to the measurement of certain peptides / proteins that can serve as evidence of allergies.
  • this aspect of the present invention is concerned with the measurement of gamma interferon. It has to be clarified how much active gamma interferon is still in the blood in order to be able to determine the amount of gamma interferon to be added from outside for a patient.
  • ⁇ EFN undifferentiated (active plus inactive degraded) ⁇ EFN (peptides, proteins) in cell cultures, cell lines from secretion assays with stimulated isolated cells. It is also known that the active ⁇ lFN (peptides, proteins) molecules relevant for patients / persons (species) are contained in their body fluids, e.g. Blood, are metabolized very quickly, are broken down and an exact allocation of the relevant active ⁇ lFN (peptide, protein) amounts is not possible with the conventional assays.
  • the invention aims to overcome the above drawbacks.
  • the invention aims to enable an exact allocation of the relevant active ⁇ IFN (peptide, protein) amounts.
  • the invention thus provides a method for determining peptides / proteins, characterized in that peptides / proteins at their C end, N end and / or their modified sites and in their entirety with specific antibodies, which are marked such that a later measurement is possible.
  • the basic idea of the invention is based on the measurement of peptides / proteins at their C-end, N-end, their modified sites, and in their entirety (comparison with conventional ⁇ IFN assays) with specific antibodies which are labeled such that a later measurement is possible.
  • the invention leads to new methods for the determination of peptides / proteins, for the detection of active ⁇ lFN (interleukins, cytokines) and their degradation products / metabolites in blood, in plasma, in serum (body) liquids, in tissues, and for the separation of active ⁇ lFN (interleukins, cytokines) and its degradation products / metabolites from these media.
  • therapies of I ranldieitssente ⁇ i / metabolic diseases urticaria, allergies, leukaemias, thromboses, atherosclerosis, rheumatism, rheumatoid arthritis, etc.
  • therapies of clinical pictures / metabolism can be achieved which are associated with the presence of active ⁇ lFN (Interleukins, cytokines) and their degradation products / metabolites are linked, developed, selected, facilitated or improved.
  • the present invention enables a determination of peptides / proteins, for the replacement of active ⁇ lFN (interleukins, cytokines) and its degradation products / metabolites in the absence of a quantity to the normal value.
  • the present invention also makes positive contributions to the development, selection, facilitation and improvement of therapies for clinical pictures / metabolic diseases which are associated with the lack of active ⁇ IFN (hiterleukins, cytokines) and their breakdown products / metabolites, and simultaneous (proportionate) evidence of the different ⁇ lFN / interleukins, cytokines (peptides / proteins) to one another and conclusions about the effect of this special molecule on the metabolism and on the triggering or therapy of derailments.
  • the invention also makes contributions to the explanation of the effect of ⁇ IFN (11-4, II's, cytokines) peptides / proteins on the therapy of various antitumor drugs / antagonist agonists (Cyt P 450 system etc.) to explain the Change in redox potential, etc.
  • the method further provides that a detection possibility (defection device) is coupled to the antibodies, which allows the amount of the antibody molecules to be determined via their concentration. le and thus to determine the quantity of molecules to be determined qualitatively and / or quantitatively.
  • a detection possibility is coupled to the antibodies, which allows the amount of the antibody molecules to be determined via their concentration. le and thus to determine the quantity of molecules to be determined qualitatively and / or quantitatively.
  • the detection or defect device is designed to duplicate or amplify the concentration to be measured. This can be further developed by either designing the detection possibility or the defect device itself to be very large and, in particular, breaking it down into smaller, preferably identical parts in such a way that their concentration can be assumed to be known and / or easily measured, or that the detection possibility or defect Device itself can be duplicated or amplified in such a way that measurement is easily possible using known methods.
  • this also relates to mass spectrometry for metals or metal determination in general, specifically in the form of a mass spectrometer and a method for metal determination with a mass spectrometer.
  • the starting point for this aspect of the invention is conventional mass spectrometry, in which an organic sample in the chamber of the mass spectrometer is bombarded with electrons or a LASER. The resulting positively charged cations are separated by a magnetic field according to mass and charge and recorded with a detector.
  • This conventional mass spectrometry relies on volatile substances.
  • the present invention provides a mass spectrometer, which is characterized in that a carrier gas source, in particular a CO source, is provided in order to bombard a sample with carrier gas, corresponding in particular CO, or that an enzyme solution container with / for an enzyme solution is provided for reaction with at least one metal.
  • a carrier gas source in particular a CO source
  • the aim of the invention is thus achieved by replacing the electron source, which is conventionally customary, for example, with a CO source or generally a carrier gas source, in particular metal-containing fluids being introduced into a sample chamber.
  • the Trä- Gas source or in particular CO source causes the metals contained in the fluids to volatilize in carbonyls and can thus be recorded by the detector.
  • Metal determination method which is characterized in that a sample is bombarded with carrier gas, in particular CO, from a carrier gas source, corresponding in particular with a CO source, or that an enzyme solution reacts with at least one sample metal.
  • Metal-containing fluids are preferably introduced into a sample chamber as part of this method.
  • Carrier gas or, in particular, CO radiation or carrier gas or, in particular, CO bombardment causes the metals contained in the fluids to evaporate in carbonyls and thus to be recorded by the detector.
  • inorganic cations Pt (2+), Pd (2+), Ni (2+), incl.
  • These metals can not or only insufficiently in low concentration with the currently common methods (AAS, etc.) be measured.
  • the measurement of these metals is particularly interesting with regard to various environmental diseases such as contact dermatitis, asthma or BSE, as well as the ozone hole and the greenhouse effect.
  • These metals usually have a low vapor pressure. The tendency of these metals to form volatile carbonyls is used, which makes these metals detectable in the mass spectrometer. Car exhaust gases can then also be measured directly, making the device and method suitable for technical inspections of motor vehicles, such as TÜV in Germany, as well as atmospheric metals and compounds (complexes) as well as organics etc. more.
  • the mass spectrometer and mass determination method according to the invention are designed to determine the concentration of the metals by determining contained and / or emerging peptides, amino acids, etc. This is preferably, but not limited to, e.g. PCR reaction or the methods described above according to the first aspect of the present invention.
  • the metals can also be determined by means of an enzymatic reaction.
  • these metals are introduced into corresponding enzyme solutions which react specifically with these metals, ie are activated (for example nickel-activated horse radical peroxidase, Me proteases, superoxide dismutase, glutathione peroxidase, etc.).
  • this also relates to conductivity measurement and conductivity determination.
  • a biosensor with two electrodes, to which a constant voltage is applied is known.
  • Blood cells for example, are placed on the electrodes, the change in resistance being measured when the cells are added to the electrodes.
  • cell activity can also be measured when the blood cells are already on the electrodes and additional substances are added.
  • this method cannot be used to measure cells that are still alive. It should be noted that cells can still be measured for about half an hour under in vivo conditions after blood has been drawn.
  • the third aspect of the present invention aims to remedy this drawback.
  • a conductivity measurement according to the invention is characterized in that a blood sample is introduced into a measuring tube, which is surrounded by a plurality of electrodes, and that the electrodes are controlled crosswise with a frequency generator according to a predetermined scheme.
  • a measuring tube is provided for the introduction of a blood sample, which is surrounded by a plurality of electrodes which can be controlled crosswise with a frequency generator according to a predefinable scheme.
  • the invention in the context of the aspect dealt with here therefore consists in introducing the blood sample into a measuring tube which is surrounded by a multiplicity of electrodes.
  • the electrodes are controlled crosswise according to a certain scheme with a frequency generator.
  • this method it is possible, for example, to measure certain redox conditions; overall, it is possible to measure the patient's individual reaction to certain additives.
  • the drawing shows: 1 shows a molecule with modification sites on the example of ⁇ IFN in the context of the method according to the invention for the determination of peptides / proteins,
  • FIG. 3 shows an exemplary embodiment of a protein, peptide or ⁇ IFN with antibody with a defect device as detection means, within the scope of the method according to the invention for determining peptides / proteins,
  • Fig. 4 shows another embodiment of a ⁇ lFN with antibody with a defect
  • FIG. 5 shows a diagrammatic representation of the separation of the defection device as detection means, within the scope of the method according to the invention for determining peptides / proteins
  • FIG. 6 shows an exemplary embodiment of a conversion step in the context of the method according to the invention for determining peptides / proteins
  • FIG. 7 shows an exemplary embodiment of a detection step in the context of the method according to the invention for determining peptides / proteins
  • FIG. 8 shows an exemplary embodiment of a calculation step within the scope of the method according to the invention for determining peptides / proteins
  • FIG. 8a shows an embodiment variant according to FIG. 8, but especially for a missing N end
  • 11 still exemplary embodiments of method variants within the scope of the method according to the invention for the determination of peptides / proteins
  • 12 shows another example of the implementation of process variants within the scope of the method according to the invention for determining peptides / proteins
  • FIG. 17 schematically shows two exemplary embodiments of a conductivity measuring cell
  • FIG. 1 shows a molecule such as ⁇ IFN (identical to the notation gIFN) with different modification sites, with 0. the entire active molecule, 1. to 3. the degradation product, th and 4th to 6th correspond to the modification points.
  • Figure 2 shows a molecule like gIFN that lacks the N-terminus.
  • a monoclonal antibody AK specific for the C-end is added to the sample. Then shake for a few minutes.
  • Antibodies AK are preferably commercially available or newly manufactured. The principle applies in particular to every protein (molecule) to be measured, modified or changed.
  • ⁇ IFN identical to the notation gIFN
  • U / ml blood 2.5-25 ng / ml blood.
  • the corresponding detection possibility or Defection Device DD is preferably able to reproduce or amplify the concentration to be measured in such a way that the measurement of the unknown concentration is easily possible, for example also in normal blood samples is.
  • the determination method for peptides / proteins can provide that the detection possibility or Defection Device DD itself is designed to be very large and in particular can be broken down into smaller, preferably identical, parts such that their concentration can be assumed to be known and / or easily measured , In another variant it can be provided that this detection possibility or Defection Device DD itself can be reproduced or amplified in such a way that measurement is easily possible using known methods, for example by means of a subsequent PCR reaction.
  • the detection possibility or defect device DD can then be a magnetic particle, an RNA, a DNA, a polylysine, a polysaccharide, a polypeptide, etc., which is coupled via PCR, magnetically, electrically, electrophoretically (blotting) Dyes (UV, IR, luminescence, fluorescence, etc.) can be easily measured qualitatively and quantitatively.
  • a coupled DNA or RNA is e.g. easy to disassemble.
  • modified ⁇ lFN e.g. commercially available poly (Ad) or poly (G; T; C) chains (balls) are suitable, which would be coupled to an antibody AK in the context of the invention using any coupling method.
  • Ad poly (Ad) or poly (G; T; C) chains (balls) are suitable, which would be coupled to an antibody AK in the context of the invention using any coupling method.
  • These coupling methods are e.g. used routinely in DNA chip technology to couple these compounds to the biochip (see Figure 4).
  • This complex can now be detected in the sample (blood etc.) directly and / or after appropriate separation or washing (centrifugation, TLC plate, column means, inter alia, polyclonal AKs against the entire molecule and digestion of the entire molecule by proteases, peptidases) ,
  • DD ⁇ Poly (Ad) ⁇ is via a chip or a microtiter plate (or other possibilities) by e.g. easily isolate or separate hybidization from (T) covered / occupied surfaces from the sample.
  • the detection is carried out radioactively using labeled nucleotides, 125 J antibodies, etc. Or the detection is carried out by means of a coupled PCR reaction or after digestion of the DNA, RNA, poly (A) (chains) by means of acid bases, DNAases, RNAases, endonucleases etc., followed by an enzyme. Detection of the released nucleotides carried out (1st digestion with common methods to monophosphates).
  • the triphosphates / ATP formed are now detected, for example, by means of downstream enzymes.
  • a measurement of the kinase activity is also possible.
  • the most sensitive method, however, is the measurement using coupled luciferase via luminescence.
  • Detection is also possible using RNA-F, DNA-F, polyNu-F (polylysine-F, polysaccharide-F, etc.) labeled with fluorescent dye (cf. FIG. 7).
  • the concentration calculation is carried out, for example, in accordance with the embodiment shown in FIG. 8 as process component I.
  • process component II is missing for the C-terminus: analogous to L, but with an antibody AK specifically for the N end according to FIG. 8a.
  • a process component HI applies. analogously I. with antibodies AK specifically for these modified molecules, etc.
  • a process component IV applies to the modified molecule ( ⁇ IFN (identical to the notation gIFN)) with policlonal antibodies AK against the modified molecule.
  • the surfaces are coupled with the corresponding counter nucleotides, so that hybridization can take place.
  • the variant a) in FIG. 10 relates to a microtiter plate, etc. for separation or as proof.
  • the variant b) in FIG. 10 relates to a chip, a tube (advantageously, for example, in dialysis) or a column, etc. for separation or for detection.
  • polypeptides are detected using common assays, such as with ninhydrin (Edman degradation, etc.).
  • Evidence of poly sugar e.g. Chitisan, done by coupling with e.g. Dyes, degradation with chitobiose-chitobiase, detection as described (chelates, flocculants for metals).
  • Another way of determining the complex is achieved by means of atomic force microscopy (AFM) titration (cf. FIG. 11).
  • AFM atomic force microscopy
  • the isolated, labeled complexes are applied directly from the separation columns, using nanopipettes, etc. to a suitable sample holder (glass, paper, nylon, etc.) in such a way that a number of spots with increasing concentration arises, which after drying (+ fixation with e.g. formalin, coupling with isothiocyanates, epoxy (-D3), NHS, etc.) are measured orthographically using a CCD camera, scintillation counter, etc.
  • cytokines for testing and control, incubate for certain time intervals and then measure the concentration of cytokines, interferons, etc. ( ⁇ lFN, interleukin-1 (ß), IGF BPs, IGFI etc.) using the methods described.
  • a combination of the different assays on a microtiter plate or on a biochip is also possible.
  • FIG. 13 shows the structure of a conventional mass spectrometer 1 with an electron source 2, an accelerator 3, in particular with a negative potential, a magnet 4, a gap 5 and a collecting vessel 6. Furthermore, a line 14 to a vacuum pump (not shown) is shown and a feeder 15 for a sample (not shown).
  • valve 14 shows the representation of a mass spectrometer analogous to FIG. 1 with the configurations and arrangements according to the invention: valve / switch 7, heatable sample space with furnace 8, (air, gas) CO source / reservoir 9, valve 10 and valve 11 (three - directional valves).
  • the electron source 2 can be switched off according to the schematic illustration in FIG. 14 by means of the valve / switch 7 and the heatable sample space with sample 8 and CO (air, gas, carrier gas) source 9 by means of valves (in particular three-way valves) 10 and 11 connectable to the mass spectrometer 1.
  • Exhaust gases are also (Air, atmosphere, car exhaust gases) can be supplied via the valve 11.
  • the heatable sample space 8 can be opened by means of a closure 12 and can be loaded with a sample carrier / sample 13.
  • the sample space 8 with the sample e.g. Blood or other metal-containing fluids can be loaded with the aid of the closure 12.
  • the CO source 9 and the sample space 8 are connected to the sample 13 via the valves 10 and 11 with the mass spectrometer 1.
  • the sample is heated in the heatable sample room (e.g. to 300 ° C), whereby volatile metal carbonyls are formed which, by means of a vacuum pump and accelerator 3, pass through the magnet 4 and the gap 5 into the collecting vessel 6 after they have been detected.
  • Exhaust gases e.g. Car exhaust gases, air, atmosphere, etc. are introduced directly into the system via valve 11 and measured / detected.
  • the detection limit is preferably in the vicinity of 0.1 pmol / ⁇ l sample.
  • 15 shows the mass spectrogram of ethanol as an example.
  • 16 shows an example of the assumed mass peak of Pt 2+.
  • the metals can also be determined by means of an enzymatic reaction.
  • these metals are introduced into corresponding enzyme solutions which react specifically with these metals, i.e. activated (e.g. nickel-activated horse radical peroxidase, Me proteases, superoxide dismutase, glutathione peroxidase, etc.).
  • activated e.g. nickel-activated horse radical peroxidase, Me proteases, superoxide dismutase, glutathione peroxidase, etc.
  • the determination of the resulting peptides, amino acids, etc. using e.g. PCR reaction or the methods described above gives the concentration of the metals.
  • the specific inactivation of various enzymes by these metals can also be used to determine the metals (and vice versa), these are proteases, peptidases, transferases, etc.
  • a microtiter plate (e.g. a chip, etc.) covered with glutathione, metallothionine, transferrin, ferritin, dextrin (or another chelating agent, fusion protein, etc.) binds e.g. Nickel from a sample (liquid, air, etc.) (1), a recombinant horse radish peroxidase is activated (2) and can be removed using substrates such as e.g. Polyhistidine and PCR (among others) detection show the concentration of nickel in the sample.
  • substrates such as e.g. Polyhistidine and PCR (among others) detection show the concentration of nickel in the sample.
  • Aeroallergens such as metal ions
  • B SA filters / columns can be enriched via B SA filters / columns and then detected in / from these filters / columns using the methods described.
  • a conductivity determiner can then also be used to determine these ions, as is explained below in connection with yet another aspect of the present invention.
  • redox potentials in biol are defined by Me ions. Systems, among others.
  • the invention in the context of this aspect consists in introducing a blood sample into a measuring tube which is surrounded by a plurality of electrodes.
  • the electrodes are controlled crosswise according to a certain scheme with a frequency generator.
  • this method it is possible, for example, to measure certain redox conditions; overall, it is possible to measure the patient's individual reaction to certain additives.
  • this redox measuring cell / conductivity measuring cell enables the "ex vivo" (in vivo) measurement of the potential over a (biological) sample, blood sample (cell (s), over cell components, membranes - mitochondria, bacteria (etc.), Cell nucleus cytosol, solutions; blood and its components / serum, plasma; etc., with and without different compounds, under different treatments, conditions.
  • FIG. 17 shows an example for the construction of a conductivity measuring cell 100 with electrodes 102, an opening 103, and feed lines 104 for a frequency generator 105 (FIG. 18).
  • the conductivity measuring cell 100 is designed in Kugelfönn (FIG. 17, example a)), one side being open for the addition of various substances, additives, etc.
  • the total internal volume of the conductivity measuring cell 100 is preferably approximately 0.5 ml.
  • Suitable electrode material would be e.g. Gold.
  • An electromagnetic rotating field, in particular constantly circulating and / or oscillating, is generated by means of the frequency generator 105 (FIG. 18).
  • the conductivity measuring cell 100 is designed in a continuous form (FIG. 17, example b)), the conductivity measuring cell 100 being open at the top and bottom for the passage of liquids (such as, for example: blood). Openings 103 and 107 are provided with a network 106 and 108.
  • the conductivity measuring cell 100 can be used as a perfusion cell.
  • At least two conductivity measuring cells 100 are preferably connected in series, at least one conductivity measuring cell 100 acting as a control.
  • a computer controls voltage / current as well as the resistance and at the same time ensures the change between the measuring cells within an experiment.
  • the frequency generator 105 works as a current source with 4 output channels (I J to L) whose current or voltage phases are each shifted by 90 ° (ij to i 4 , with Oi to O 4 , 11), which leads to a rotating field that serves to stabilize the sample.
  • an amplifier 109 for tapping the voltage Uj at the frequency generator 105 is interposed in front of the computer 110.
  • the behavior (redox, etc.) of an individual species in the measuring cell can be measured with a known cell / particle number in the conductivity measuring cell 100.
  • redox potentials such as Fe 2 Fe 3+ ; ascorbate; glutathione, amino acids, etc.
  • cell biosystems including prion proteins, depending on various conditions (reference to BSE, Kreutzfeld-Jakob), from allergies, asthma in relation to the addition / effect of metals, ROS.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention concerne un procédé de détermination des peptides/protéines, caractérisé en ce que les peptides/protéines sont mesurés au niveau de leurs extrémités C, de leurs extrémités N et/ou de leurs points modifiés, en plus de leur totalité, en utilisant des anticorps spécifiques qui sont marqués de telle façon qu'une mesure ultérieure soit possible. L'invention concerne en outre un procédé et un dispositif de spectrométrie de masse consistant à utiliser une source de gaz porteur, en particulier une source de CO, en vue de bombarder un échantillon avec un gaz porteur, notamment CO, ou encore à utiliser un récipient de solution enzymatique avec/pour une solution d'enzyme en vue de sa réaction avec au moins un métal. L'invention concerne en outre un procédé et un dispositif permettant une mesure de conductivité, selon lequel un échantillon sanguin est introduit dans un tube de mesure qui est entouré d'une pluralité d'électrodes, lesdites électrodes étant contrôlées par croisement, conformément à un schéma prédéterminé, au moyen d'un générateur de fréquence.
EP02750770A 2001-05-29 2002-05-28 Procede de determination de peptides/proteines, spectrometrie de masse pour metaux et determination de la conductivite Withdrawn EP1397687A2 (fr)

Applications Claiming Priority (7)

Application Number Priority Date Filing Date Title
DE10126696 2001-05-29
DE10126695 2001-05-29
DE10126695 2001-05-29
DE2001126697 DE10126697A1 (de) 2001-05-29 2001-05-29 Proteinbestimmung
DE10126697 2001-05-29
DE10126696 2001-05-29
PCT/DE2002/001966 WO2002097440A2 (fr) 2001-05-29 2002-05-28 Procede de determination de peptides/proteines, spectrometrie de masse pour metaux et determination de la conductivite

Publications (1)

Publication Number Publication Date
EP1397687A2 true EP1397687A2 (fr) 2004-03-17

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EP02750770A Withdrawn EP1397687A2 (fr) 2001-05-29 2002-05-28 Procede de determination de peptides/proteines, spectrometrie de masse pour metaux et determination de la conductivite

Country Status (3)

Country Link
EP (1) EP1397687A2 (fr)
AU (1) AU2002344245A1 (fr)
WO (1) WO2002097440A2 (fr)

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4666865A (en) * 1984-01-13 1987-05-19 Centocor, Inc. Immunoassay for biologically active human interferon-gamma employing unique monoclonal antibodies
US4792521A (en) * 1985-08-15 1988-12-20 Immunomedics, Inc. Non-enzymatic immunohistochemical staining system and reagents
EP0689606A1 (fr) * 1993-03-19 1996-01-03 The Johns Hopkins University ANTICORPS ET TITRAGES PERMETTANT DE DETERMINER DES MUTATIONS DANS LE GENE $i(APC)
DE19918141A1 (de) * 1999-04-21 2000-10-26 Boehringer Ingelheim Vetmed Verfahren zur Diagnose von übertragbaren Spongiformen Enzephalopathien
US6362317B1 (en) * 1999-07-12 2002-03-26 The United States Of America As Represented By The Department Of Health And Human Services Anti-γ-H2A antibody, fusion proteins thereof and method and kit for determining DNA double-stranded breaks

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
None *
See also references of WO02097440A3 *

Also Published As

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WO2002097440A2 (fr) 2002-12-05
WO2002097440A3 (fr) 2003-11-06
AU2002344245A1 (en) 2002-12-09

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