[go: up one dir, main page]

WO2005110955A1 - Acetophenones substituees utilisees comme agents de spectrometrie de masse - Google Patents

Acetophenones substituees utilisees comme agents de spectrometrie de masse Download PDF

Info

Publication number
WO2005110955A1
WO2005110955A1 PCT/SE2005/000677 SE2005000677W WO2005110955A1 WO 2005110955 A1 WO2005110955 A1 WO 2005110955A1 SE 2005000677 W SE2005000677 W SE 2005000677W WO 2005110955 A1 WO2005110955 A1 WO 2005110955A1
Authority
WO
WIPO (PCT)
Prior art keywords
reagent
bromoacetophenone
reagents
labelled
acid
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.)
Ceased
Application number
PCT/SE2005/000677
Other languages
English (en)
Inventor
Henrik Neu
Lee Olsen
Gavin E. Reid
Kade D. Roberts
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.)
Ludwig Institute for Cancer Research Ltd
Cytiva Sweden AB
Global Life Sciences Solutions USA LLC
Ludwig Cancer Research
Original Assignee
Ludwig Institute for Cancer Research Ltd
Amersham Bioscience AB
GE Healthcare Bio Sciences Corp
Ludwig Cancer Research
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 Ludwig Institute for Cancer Research Ltd, Amersham Bioscience AB, GE Healthcare Bio Sciences Corp, Ludwig Cancer Research filed Critical Ludwig Institute for Cancer Research Ltd
Publication of WO2005110955A1 publication Critical patent/WO2005110955A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C49/00—Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
    • C07C49/76—Ketones containing a keto group bound to a six-membered aromatic ring
    • C07C49/80—Ketones containing a keto group bound to a six-membered aromatic ring containing halogen
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C309/00—Sulfonic acids; Halides, esters, or anhydrides thereof
    • C07C309/01—Sulfonic acids
    • C07C309/28—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
    • C07C309/44—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton containing doubly-bound oxygen atoms bound to the carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C317/00—Sulfones; Sulfoxides
    • C07C317/24—Sulfones; Sulfoxides having sulfone or sulfoxide groups and doubly-bound oxygen atoms bound to the same carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C317/00—Sulfones; Sulfoxides
    • C07C317/44—Sulfones; Sulfoxides having sulfone or sulfoxide groups and carboxyl groups bound to the same carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
    • C07C45/45—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds by condensation
    • C07C45/46—Friedel-Crafts reactions
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/05—Isotopically modified compounds, e.g. labelled
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2458/00—Labels used in chemical analysis of biological material
    • G01N2458/15—Non-radioactive isotope labels, e.g. for detection by mass spectrometry

Definitions

  • MS mass spectrometry
  • proteomics The field of research known as proteomics involves the systematic identification of all proteins expressed by a particular cell or tissue type at a given time, quantitative analysis of the differences in protein expression observed between two different cell states, (e.g. normal versus diseased), and identification and characterization of the protein modifications, protein complexes and specific protein-protein interactions involved in the control of cellular function.
  • MS biological mass spectrometry
  • MS/MS tandem mass spectrometry
  • Identification of these selected peptides is then performed by either database analysis of the uninterpreted product ion spectrum, through database searching of a partially derived amino acid "sequence tag", or by "de-novo" sequence analysis.
  • One of the limitations of this general analysis strategy is that typically, peptides observed at high relative abundance are preferentially selected for analysis, such that information regarding the identity of proteins represented as low abundance peptides is commonly not obtained.
  • a fundamental aspect of proteomics research is the determination of protein expression levels between two different states of a biological system (i.e., relative quantification of protein levels), such as that encountered between a normal and diseased cell or tissue.
  • relative quantification of protein levels i.e., relative quantification of protein levels
  • One approach involves the in vivo metabolic incorporation of isotopically depleted ( 13 C-, 15 N-, and 2 H-depleted) or enriched amino acids (either by uniform labelling with 15 N, or by incorporation of selected amino acids containing heavy isotopes (eg., 13 C, 15 N, 2 H)) into a cellular protein population.
  • isotopically depleted 13 C-, 15 N-, and 2 H-depleted
  • enriched amino acids either by uniform labelling with 15 N, or by incorporation of selected amino acids containing heavy isotopes (eg., 13 C, 15 N, 2 H)
  • the sample is combined with one incorporating natural isotopes, then the combined protein mixture is subjected to proteolysis and the masses of the peptides determined by MS as described above.
  • cysteine containing proteins from two different cell/tissue states are reduced and S-alkylated with either naturally abundant (light) or isotopically enriched (heavy) ICAT reagents, respectively, each containing a biotin moiety for subsequent affinity selection of cysteine-containing peptides following proteolysis by streptavidin affinity purification, leading to simplification of the mixture prior to MS analysis.
  • the present inventors have developed novel reagents for the selective identification and differential quantitation of peptides containing selected amino acids, using tandem mass spectrometry, having an increase in selectively and sensitivity of several orders of magnitude over existing reagents.
  • the present invention provides reagents for introducing a fixed charge to a protein/peptide.
  • the fixed-charge may be contained on the side-chain of a selected amino acid, or the side-chain of a selected amino acid residue contained within a protein or peptide.
  • the selected amino acid residue is methionine.
  • the fixed-charge includes a sulfonium ion.
  • the present invention is based on the fixed charge derivative contained within a peptide ion directing the MS/MS dissociation of peptides containing the fixed charge toward a single predictable fragmentation channel with the formation of a single product ion characteristic of fragmentation occurring at the fixed-charge site, thereby allowing its selective identification from a complex mixture without need for prior resolution or otherwise enrichment of the peptide from a complex mixture prior to its analysis.
  • tandem mass spectrometry Analysis of derivatized proteins/peptides may be performed by tandem mass spectrometry.
  • the tandem mass spectrometer may be equipped with electrospray ionization (ESI) or matrix assisted laser desorption ionization (MALDI) interfaces to transfer the protein or peptide ion from solution into the gas-phase.
  • ESI electrospray ionization
  • MALDI matrix assisted laser desorption ionization
  • the reagents of the present invention may be used for the selective identification of peptides.
  • the reagents of the present invention may be used for the differential quantitation of peptide concentrations between two different samples, by incorporation of stable isotope labels to the reagent, to yield a 'light' form (containing only natural isotopes), and a 'heavy' form (containing isotopic or structural labels incorporated into the substituent).
  • the present inventors have discovered that a mass difference of at least 8 Daltons between 'light' and 'heavy' reagents is desired to enable sensitive and accurate differential quantitative analysis.
  • the general form of the reagents of the present invention allows a mass difference of at least 8 Daltons and up to 13 Daltons between 'light' and 'heavy' reagents.
  • the present invention relates to use of substituted acetophenone reagents in MS analysis, especially comparative MS analysis.
  • the MS analysis is MS/MS analysis.
  • the invention relates to a substituted acetophenone, or a salt thereof, or a solvate thereof, having the following formula:
  • X is CI, Br, I, sulfonic esters, perchlorate esters and chlorosulfonates
  • A is H or 2 H (D);
  • R is H, 2 U (D), 13 CH 3 , 13 CH 2 13 CH 3 , 13 CH 2 13 CH 2 13 CH 3 , 13 CH 2 13 CO 2 H, 13 CH 2 13 CH 2 13 CO 2 H, O 13 CH 3 , O 13 CH 2 13 CH 3 , O 13 CH 2 13 CO 2 H, O 13 CH 2 13 CH 2 13 CO 2 H, SO 2 13 CH 3 , SO 2 13 CH 2 13 CH 3 , SO 2 13 CH 2 13 CH 2 13 CH 3 , SO 2 13 CH 2 13 CO 2 H, SO 2 13 CH 2 13 CH 2 13 CO 2 H, or SO 3 H.
  • the invention also relates to novel acetophenones having the formula above, and wherein
  • X is CI, Br, I, sulfonic esters, perchlorate esters and chlorosulfonates,
  • A is H
  • R 9 is SO 2 CH 2 CO 2 H or SO 2 CH 2 CH 2 CO 2 H.
  • a preferred substituted acetophenone has the following formula 13 C 8 - bromoacetophenone
  • this substituted acetophenone gives a mass difference of 8 Daltons compared to the normal isotopic variant.
  • Another preferred substituted acetophenone is one according to the above formula, wherein the hydrogens on Ri are substituted for H (D).
  • this substituted acetophenone gives a mass difference of 10 Daltons compared to the normal isotopic variant.
  • a further preferred substituted acetophenone has the following formula O H
  • this substituted acetophenone gives a mass difference of 9 Daltons compared to the normal isotopic variant.
  • Yet another further preferred substituted acetophenone is one according to the above formula, wherein the hydrogens on Ri are substituted for 2 H (D).
  • This substituted acetophenone reagent gives a mass difference of 10 Daltons compared to the normal isotopic variant.
  • Yet another further preferred substituted acetophenone is one according to the above formula, wherein the hydrogens on Ri are substituted for 2 H (D).
  • This substituted acetophenone reagent gives a mass difference of 12 Daltons compared to the normal isotopic variant.
  • a further preferred substituted acetophenone reagent has the following formula 13 C ⁇ o - jt?-propyl-bromoacetophenone
  • This substituted acetophenone reagent gives a mass difference of 11 Daltons compared to the normal isotopic variant.
  • Yet another further preferred substituted acetophenone is one according to the above formula, wherein the hydrogens on Ri are substituted for H (D).
  • This substituted acetophenone reagent gives a mass difference of 13 Daltons compared to the normal isotopic variant.
  • the invention provides MS reagents comprising substituted acetophenones as described above.
  • the invention relates to use of these acetophenones as reagents, particularly MS reagents.
  • Preferred MS-reagents are I3 C 8 - bromoacetophenone, D2, 3 C 8 - bromoacetophenone and 13 C 9 -j?-methyl-bromoacetophenone.
  • the reagents of the invention may be used for selective identification of proteins/peptides.
  • the acetophenones are used for selective identification of proteins/peptides, preferably the corresponding acetophenones with natural isotopes are used for selective identification of proteins/peptides.
  • the above acetophenones with stable isotopes are used in combination with the corresponding acetophenones with natural isotopes for quantitative determination of proteins/peptides.
  • any of the substituted acetophenones mentioned above with heavy stable isotopes can be used as heavy reagents and the corresponding acetophenones with natural isotopes as light reagents.
  • a preferred kit comprises the heavy stable isotope labelled reagent C 8 bromoacetophenone and the light reagent C 8 bromoacetophenone.
  • Another preferred kit comprises the heavy stable isotope labelled reagent D 2 , C 8 bromoacetophenone and the light reagent C 8 bromoacetophenone.
  • a further preferred kit comprises the heavy reagent C 9 - jt?-methyl-bromoacetophenone and the light reagent C 9 - ?-methyl-bromoacetophenone.
  • Yet another further preferred kit comprises the heavy reagent D 2 , C 9 - jo-methyl- bromoacetophenone and the light reagent C 9 -p-methyl-bromoacetophenone. Yet another further preferred kit comprises the heavy reagent 13 C 10 - p-ethyl- bromoacetophenone and the light reagent Cio- ⁇ -ethyl-bromoacetophenone.
  • Yet another further preferred kit comprises the heavy reagent D 2 , 13 C 10 - p-ethyl- bromoacetophenone and the light reagent C ⁇ 0 -/>-ethyl-bromoacetophenone.
  • the kit may further comprise one or more containers containing: cysteine disulfide reducing agents, cysteine alkylating reagents, proteases or chemical cleavage agents, and solvents.
  • the cysteine d isulfide reducing may be: dithiothreitol (DTT), ⁇ -mercaptoethanol, tris- carboxyethyl phosphine (TCEP), and/or tributylphosphine (TBP).
  • DTT dithiothreitol
  • TCEP tris- carboxyethyl phosphine
  • TBP tributylphosphine
  • the cysteine alkylating reagents may be alkylhalides (e.g. iodoacetic acid, iodoacetamide), vinylpyridine, acrylamide or disulphide-comprising compounds, such as bis-(2-hydroxyethyl) disulphide; bis-(2-hydroxypropyl) disulphide; 3-3-dipropionamidedisulphide; 2-2'- dipyridyldisulphide.
  • reagent is DeStreakTM.
  • proteases or chemical cleavage agents may be trypsin, Endoprotemase Lys-C, Endoprotemase Asp-N, Endoprotemase Glu-C, pepsin, papain, thermolysin, cyanogen bromide, hydroxylamine hydrochloride, 2-[2'-nitrophenylsulfenyl]-3-methyl-3'-bromoindole (BNPS- skatole), iodosobenzoic acid, pentafluoropropionic acid and/or dilute hydrochloric acid.
  • the solvents may be urea, guanidine hydrochloride, acetonitrile, methanol and/or water.
  • 13 C 8 - bromoacetophenone A non-limiting method for the synthesis of one this derivative may be performed using methods known to those skilled in the art via Friedel-Crafts acylation of 13 C 6 labelled benzene with C 2 labelled bromoacetyl bromide, prepared from C 2 labelled bromoacetic acid, to yield C 8 labelled bromoacetophenone.
  • a non-limiting method for the synthesis of this derivative may be performed using methods known to those skilled in the art via Friedel-Crafts acylation of 13 C 7 labelled toluene with 13 C2 labelled bromoacetyl bromide, prepared from 13 C 2 labelled bromoacetic acid, to yield C 9 labelled -methyl-bromoacetophenone.
  • a non-limiting method for the synthesis of this derivative may be performed using methods known to those skilled in the art via Friedel-Crafts acylation of 13 C 6 labelled benzene with 13 C 2 labelled acetylchloride or 13 C 3 labelled propanoyl chloride (formed by reaction of 13 C 3 propionic acid with thionyl chloride) to yield 13 C 8 labelled acetophenone or 13 C 9 labelled propiophenone, followed by reduction [Ishimoto, K., Mitoma, Y., Nagashima, S., Tashiro, H., Prakash, G.K.S., Olah, G.A. and Tashiro, M. Chem. Comm.
  • Synthesis of this derivative may be performed using methods known to those skilled in the art, via Friedel-Crafts acylation of 13 C 6 labelled benzene with 13 C 2 labelled acetylchloride or 13 C 3 labelled propanoyl chloride (formed by reaction of 13 C 3 propionic acid with thionyl (or oxalyl, phtaloyl) chloride) to yield 13 C 8 labelled acetophenone or 13 C 9 labelled propiophenone, followed by reduction to yield 13 C 8 labelled ethylbenzene or 13 C 9 labelled propylbenzene, then Friedel-Crafts acylation with 13 C 2 , D 2 labelled acetylbromide, prepared from 13 C 2 , D 4 acetic acid or 13 C 2 , D 3 acetic acid sodium salt, to yield D 2 , C 10 labelled -ethyl-acetophenone or D2, C ⁇ labelled ?-propyl-aceto
  • a non-limiting method for the synthesis of these derivatives may be performed using methods known to those skilled in the art via oxidation [Reitsema, R.H. and Allphin, N.L. J. Org. Chem. 1962, 27, 27-28.] of 13 C 8 labelled ethylbenzene or 13 C 9 labelled propylbenzene to yield the 13 C 8 labelled phenylacetic acid or 13 C 9 labelled phenylpropionic acid, followed by Friedel-Crafts acylation with 13 C 2 labelled bromoacetyl bromide, prepared from 13 C 2 labelled bromoacetic acid, to yield 13 C 10 labelled -bromoacetyl-phenylacetic acid or 13 C ⁇ p- bromoacetyl-phenylpropionic acid.
  • a non-limiting method for the synthesis of these derivatives may be performed using methods known to those skilled in the art via nucleophilic substitution of a 13 C l5 13 C2, or 13 C 3 labelled haloalkane or haloalkanoic acid with 13 C 6 labelled phenol, followed by Friedel-Crafts acylation [Earle, M.J., Seddon, K.R., Adams, CJ. and Roberts, G. Chem. Comm.
  • a non- limiting method for the synthesis of these derivatives may be performed using methods known to those skilled in the art via reaction of 13 C 6 labelled bromobenzene with 13 C ⁇ , 13 C 2 or 13 C 3 labelled sodium alkylthiolate [Shaw, J. E. J. Org. Chem. 1991, 56, 3728-3729.], followed by Friedel-Crafts acylation [Abdur Rahim, M., Praveen Rao, P.N., and Knaus, E.E., Bioorg. Med. Chem. Lett. 2002, 12, 2753-2756.; Cutler, R.A., Stenger, R.J., Suter, CM. J. Am. Chem. Soc.
  • Benzenethiol or 13 C 6 - labelled benzenethiol is then reacted with a haloalkanoic acid or 13 C l5 13 C 2 or 13 C 3 labelled haloalkanoic acid, followed by Friedel-Crafts acylation [Walker, D. and Leib, J. J. Org. Chem. 1963, 28, 3077-82.] with acetylchloride or 13 C 2 labelled acetylchloride, then oxidation [Webb, K. Tet. Lett.
  • Acetyl chloride (0.0264mol, 1.86 mL) was slowly added to a mixture of Aluminium chloride (0.0901, 12g) in carbon disulfide (40 mL) with stirring and cooling in an ice/water bath, which was followed by the gradual addition of Phenylthiopropionic acid. The mixture became quite thick and difficult to stir so Nitrobenzene (10 mL) was added to help ease the stirring. The reaction mixture was left to stir overnight ( ⁇ 16 hrs) at room temp. The reaction was quenched (destroy excess A1C1 3 ) by the slow addition of 2.5M HC1 (70-80 mL) with cooling in an ice/water bath.
  • reaction solution was acidified ( ⁇ pH 2) with cone HC1.
  • THF (20 mL) was added to redissolve the precipitate, followed by the addition of Ethyl acetate (30 mL).
  • the organic layer was separated, washed with water (1 x 40 mL) then dried over Na 2 SO 4 .
  • the solvent was removed invacuo to yield 4- Acetylphenylsulfonylpropionic acid (1.7g, 74%) as an off white solid.
  • Polyvinylpyridinium tribromide resin was added to a solution of 4- Acetylphenylsulfonylpropionic acid (0.00196 mol, 0.5g) in THF (20 mL). The resulting mixture was stirred at room temp overnight ( ⁇ 16 hrs). The mixture was filtered and the THF removed at atmosphere under a stream of nitrogen. The bright orange residue was dissolved in ethyl acetate (20 mL), which was then washed with water (1 x 20 mL) and dried over Na 2 SO 4 . The Ethyl acetate was removed at atmosphere under a stream of nitrogen.
  • a non- limiting method for the synthesis of these derivatives may be obtained using methods known to those skilled in the art via reaction of 13 C 6 benzene with 2,4- dinitrophenylsulfenyl chloride [Buess, CM. and Kharasch, N. J. Am. Chem. Soc. 1950, 72, 3529-3532.; Kharasch, N. and Swidler, R. J. Org. Chem. 1954, 19, 1704-1707] to yield 13 C 6 - phenyl 2,4-dinitrophenyl sulfide, followed by Friedel-Crafts acylation [Szmant, H.H. and Irwin, D.A. J Am. Chem. Soc.
  • the reagents of the invention are intended for the side chain fixed-charge derivatization of peptides containing methionine to enable selective identification and differential quantitation of peptides by directed and selective fragmentation during MS/MS dissociation.
  • the rationale for using fixed-charge derivatives of methionine containing peptides is based on the idea that approaches for selective protein identification and differential quantitation of protein expression should be specific for the detection of peptides containing amino acids that are rare, thereby limiting the number of peptides required to be analyzed, yet providing comprehensive coverage of proteins in the sequence databases.
  • the improved reagents according to the invention give increased mass differences compared to known reagents. This is especially useful for differential quantification between two samples.
  • the increased mass difference provides higher sensitivity and enables improved analysis of low abundant proteins/peptides.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)

Abstract

L'invention concerne des réactifs et des trousses permettant d'introduire une charge fixe dans la chaîne latérale de la méthionine, ainsi que des peptides et des protéines contenant de la méthionine et permettant une identification sélective par spectrométrie de masse tandem. Les réactifs sont basés sur des acétophénones substituées représentées par la formule (I). Les réactifs de la présente invention peuvent également être utilisés pour la quantification différentielle de concentrations peptidiques entre deux échantillons différents par incorporation de marqueurs isotopiques stables dans le réactif en vue de l'obtention d'une forme 'légère' (contenant uniquement des isotopes naturels), une forme 'lourde' étant obtenue par incorporation de marqueurs isotopiques stables dans le substituant. Ces réactifs permettent d'obtenir une différence de masse comprise entre au moins 8 daltons et 13 daltons entre les réactifs 'légers' et 'lourds'.
PCT/SE2005/000677 2004-05-17 2005-05-10 Acetophenones substituees utilisees comme agents de spectrometrie de masse Ceased WO2005110955A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE0401293A SE0401293D0 (sv) 2004-05-17 2004-05-17 Novel MS Reagents
SE0401293-6 2004-05-17

Publications (1)

Publication Number Publication Date
WO2005110955A1 true WO2005110955A1 (fr) 2005-11-24

Family

ID=32501931

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SE2005/000677 Ceased WO2005110955A1 (fr) 2004-05-17 2005-05-10 Acetophenones substituees utilisees comme agents de spectrometrie de masse

Country Status (2)

Country Link
SE (1) SE0401293D0 (fr)
WO (1) WO2005110955A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9202678B2 (en) 2008-11-14 2015-12-01 Board Of Trustees Of Michigan State University Ultrafast laser system for biological mass spectrometry
CN105699580A (zh) * 2016-04-28 2016-06-22 河南大学 基于柱前衍生的lc-ms测定有机酸含量的方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003027682A2 (fr) * 2001-09-27 2003-04-03 Purdue Research Foundation Materiaux et procedes regulant les effets isotopiques lors du fractionnement d'analytes
WO2004046731A2 (fr) * 2002-11-18 2004-06-03 Ludwig Institute For Cancer Research Methode permettant d'analyser des acides amines, des peptides et des proteines

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003027682A2 (fr) * 2001-09-27 2003-04-03 Purdue Research Foundation Materiaux et procedes regulant les effets isotopiques lors du fractionnement d'analytes
WO2004046731A2 (fr) * 2002-11-18 2004-06-03 Ludwig Institute For Cancer Research Methode permettant d'analyser des acides amines, des peptides et des proteines

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9202678B2 (en) 2008-11-14 2015-12-01 Board Of Trustees Of Michigan State University Ultrafast laser system for biological mass spectrometry
CN105699580A (zh) * 2016-04-28 2016-06-22 河南大学 基于柱前衍生的lc-ms测定有机酸含量的方法

Also Published As

Publication number Publication date
SE0401293D0 (sv) 2004-05-17

Similar Documents

Publication Publication Date Title
JP4188701B2 (ja) タンパク質サンプルを分析するためのプロセス
CN103392219B (zh) 通过质谱法定量胰岛素
US20080242838A1 (en) Labeling reagent and methods of use
Bollineni et al. Qualitative and quantitative evaluation of derivatization reagents for different types of protein-bound carbonyl groups
US11105810B2 (en) Reagents for quantitative mass spectrometry
EP2087357A1 (fr) Composés et procédés de double marquage de polypeptides pour permettre le multiplexage en analyse spectrométrique de masse
US20060094121A1 (en) Method for analysing amino acids, peptides and proteins
Zaia et al. Comparison of charged derivatives for high energy collision-induced dissociation tandem mass spectrometry
EP1565752A2 (fr) Methode permettant d'analyser des acides amines, des peptides et des proteines parspectrometrie de masse de leurs derives modifies par des charges fixees
Fenn et al. Enhanced carbohydrate structural selectivity in ion mobility-mass spectrometry analyses by boronic acid derivatization
CA2490271A1 (fr) Procede et reactif destines a l'identification specifique et a la quantification d'au moins une proteine dans un echantillon
WO2005110955A1 (fr) Acetophenones substituees utilisees comme agents de spectrometrie de masse
US20050042676A1 (en) Characterising polypeptides
JP2004532419A (ja) ポリペプチドの特徴分析方法
EP3155436B1 (fr) Marqueurs de masse isobariques
US20090053817A1 (en) Fixed charge reagents
EP1763510B1 (fr) Procede de sequençage et de quantification d'acides amines dans des polypeptides
EP2710380B1 (fr) Procédé de détection de sequences d'acides amines et/ou identification de peptides et de protéines par utilisation d'un réactif de dérivatisation et synthese d'acide 5-formyle benzo-1,3-disulphonique comme réactif de dérivatisation
JP2010509569A (ja) タンパク質ラベリングにおけるアリールボロン酸の使用
WO2006023398A2 (fr) Reactifs de spectrometrie de masse marques par des isotopes modulaires et procedes d'analyse quantitative d'acides amines, de peptides et de proteines
Kurono et al. Quantitative protein analysis using 13C7-labeled iodoacetanilide and d5-labeled N-ethylmaleimide by nano liquid chromatography/nanoelectrospray ionization ion trap mass spectrometry
Romijn et al. Mass Spectrometry: Proteomics
WO2003102592A2 (fr) Liaison selective et analyse de macromolecules
Nakagawa et al. Modification of cysteine residue in peptides by a fluorescent reagent for complete sequence analyses using post-source decay method in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
AU2002310610A1 (en) Characterising polypeptides

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KM KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NG NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SM SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LT LU MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DPEN Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed from 20040101)
NENP Non-entry into the national phase

Ref country code: DE

WWW Wipo information: withdrawn in national office

Country of ref document: DE

122 Ep: pct application non-entry in european phase