Title:
INVERSE LABELING METHOD FOR THE RAPID IDENTIFICATION OF MARKER/TARGET PROTEINS
Document Type and Number:
WIPO Patent Application WO2002052271
Kind Code:
A3
Abstract:
A novel procedure for performing protein labeling for comparative proteomics termed inverse labeling is provided for the rapid identification of marker or target proteins. With this method, to evaluate protein expression of a disease or a drug treated sample in comparison with a control sample, two converse collaborative labeling experiments are performed in parallel. In one experiment the perturbed sample (by disease or by drug treatment) is isotopically heavy-labeled, whereas, the control is isotopically heavy-labeled in the second experiment. Whem mixed and analyzed with its unlabeled or isotope light counterpart for differential comparison, a characteristic inverse labeling pattern is observed between the two parallel analyses for proteins that are differentially expressed to an apreciable level. In particularly useful embodiments, protein labeling is achieved through proteolytic <18>O-incororation into peptides as a result of proteolysis performed in <18>O-water, metabolic incoroporation of <15>N(or <13>C and <2>H) into proteins, and chemically tagging proteins with an isotope-coded tag reagent such as an isotope-coded affinity tag reagent.
Inventors:
WANG YINGQI KAREN (US)
MA ZHIXIANG (US)
QUINN DOUGLAS FREDERICK (US)
FU EMIL W (US)
MA ZHIXIANG (US)
QUINN DOUGLAS FREDERICK (US)
FU EMIL W (US)
Application Number:
PCT/EP2001/015228
Publication Date:
October 31, 2002
Filing Date:
December 21, 2001
Export Citation:
Assignee:
NOVARTIS AG (CH)
NOVARTIS ERFIND VERWALT GMBH (AT)
WANG YINGQI KAREN (US)
MA ZHIXIANG (US)
QUINN DOUGLAS FREDERICK (US)
FU EMIL W (US)
NOVARTIS ERFIND VERWALT GMBH (AT)
WANG YINGQI KAREN (US)
MA ZHIXIANG (US)
QUINN DOUGLAS FREDERICK (US)
FU EMIL W (US)
International Classes:
G01N30/88; G01N27/62; G01N33/483; G01N33/58; G01N33/68; (IPC1-7): G01N33/68
Domestic Patent References:
| WO2001094935A2 | 2001-12-13 |
Other References:
ROSE, K. ET AL: "A new mass-spectrometric C-terminal sequencing technique finds a similarity between gamma-interferon and alfa2-interferon and identifies a proteolytically clipped gamma-interferon that retains full antiviral activity", THE BIOCHEMICAL JOURNAL, vol. 215, no. 2, 1 November 1983 (1983-11-01), pages 273 - 277, XP001079683
ODA, Y. ET AL: "Accurate quantitation of protein expression and site-specific phosphorylation", PROC. NATL. ACAD. SCI., no. 96, 1999, pages 6591 - 6596, XP001080471
JI, J. ET AL: "Strategy for qualitative and quantitative analysis in proteomics based on signature peptides", JOURNAL OF CHROMATOGRAPHY, no. 745, 2000, pages 197 - 210, XP004215299
CHEN, X. ET AL: "Site-specific mass tagging with stable isotopes in proteins for accurate and efficient protein identification", ANAL. CHEM., vol. 72, no. 6, 15 March 2000 (2000-03-15), pages 1134 - 1143, XP002207456
WANG, Y.K. ET AL: "Inverse 18 O labeling mass spectrometry for the rapid identification of marker/target proteins", ANALYTICAL CHEMISTRY, vol. 73, no. 15, 1 August 2001 (2001-08-01), USA, pages 3742 - 3750, XP001061446
ODA, Y. ET AL: "Accurate quantitation of protein expression and site-specific phosphorylation", PROC. NATL. ACAD. SCI., no. 96, 1999, pages 6591 - 6596, XP001080471
JI, J. ET AL: "Strategy for qualitative and quantitative analysis in proteomics based on signature peptides", JOURNAL OF CHROMATOGRAPHY, no. 745, 2000, pages 197 - 210, XP004215299
CHEN, X. ET AL: "Site-specific mass tagging with stable isotopes in proteins for accurate and efficient protein identification", ANAL. CHEM., vol. 72, no. 6, 15 March 2000 (2000-03-15), pages 1134 - 1143, XP002207456
WANG, Y.K. ET AL: "Inverse 18 O labeling mass spectrometry for the rapid identification of marker/target proteins", ANALYTICAL CHEMISTRY, vol. 73, no. 15, 1 August 2001 (2001-08-01), USA, pages 3742 - 3750, XP001061446
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