PMID- 16112985
OWN - NLM
STAT- MEDLINE
DCOM- 20060328
LR  - 20210217
IS  - 1535-9476 (Print)
IS  - 1535-9476 (Linking)
VI  - 4
IP  - 12
DP  - 2005 Dec
TI  - Rapid and sensitive identification of major histocompatibility complex class 
      I-associated tumor peptides by Nano-LC MALDI MS/MS.
PG  - 1888-97
AB  - Identification of major histocompatibility complex (MHC)-associated peptides 
      recognized by T-lymphocytes is a crucial prerequisite for the detection and 
      manipulation of specific immune responses in cancer, viral infections, and 
      autoimmune diseases. Unfortunately immunogenic peptides are less abundant species 
      present in highly complex mixtures of MHC-extracted material. Most peptide 
      identification strategies use microcapillary LC coupled to nano-ESI MS/MS in a 
      challenging on-line approach. Alternatively MALDI PSD analysis has been applied 
      for this purpose. We report here on the first off-line combination of nanoscale 
      (nano) LC and MALDI TOF/TOF MS/MS for the identification of naturally processed 
      MHC peptide ligands. These peptides were acid-eluted from human leukocyte antigen 
      (HLA)-A2, HLA-A3, and HLA-B/-C complexes separately isolated from a renal cell 
      carcinoma cell lysate using HLA allele-specific antibodies. After reversed-phase 
      HPLC, peptides were further fractionated via nano-LC. This additional separation 
      step provided a substantial increase in the number of detectable candidate 
      species within the complex peptide pools. MALDI MS/MS analysis on 
      nano-LC-separated material was then sufficiently sensitive to rapidly identify 
      more than 30 novel HLA-presented peptide ligands. Peptide sequences contained 
      perfect anchor amino acid residues described previously for HLA-A2, HLA-A3, and 
      HLA-B7. The most promising candidate for a T-cell epitope is an HLA-B7-binding 
      nonamer peptide derived from the tumor-associated gene NY-BR-16. To demonstrate 
      the sensitivity of our approach we characterized peptides binding to HLA-C 
      molecules that are usually expressed at the cell surface at approximately only 
      10% the levels of HLA-A or HLA-B. In fact, multiple renal cell carcinoma peptides 
      were identified that contained anchor amino acid residues of HLA-Cw5 and HLA-Cw7. 
      We conclude that the nano-LC MALDI MS/MS approach is a sensitive tool for the 
      rapid and automated identification of MHC-associated tumor peptides.
FAU - Hofmann, Sandra
AU  - Hofmann S
AD  - Institute for Pharmaceutical Chemistry/Center for Drug Research, Development and 
      Safety (ZAFES), Biocenter, Johann Wolfgang Goethe-University of Frankfurt, 
      Marie-Curie-Str. 9, 60439 Frankfurt/Main, Germany. hofmann@iachem.de
FAU - Gluckmann, Matthias
AU  - Gluckmann M
FAU - Kausche, Sandra
AU  - Kausche S
FAU - Schmidt, Andrea
AU  - Schmidt A
FAU - Corvey, Carsten
AU  - Corvey C
FAU - Lichtenfels, Rudolf
AU  - Lichtenfels R
FAU - Huber, Christoph
AU  - Huber C
FAU - Albrecht, Christian
AU  - Albrecht C
FAU - Karas, Michael
AU  - Karas M
FAU - Herr, Wolfgang
AU  - Herr W
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20050819
PL  - United States
TA  - Mol Cell Proteomics
JT  - Molecular & cellular proteomics : MCP
JID - 101125647
RN  - 0 (Histocompatibility Antigens Class I)
RN  - 0 (Neoplasm Proteins)
RN  - 0 (Peptide Fragments)
SB  - IM
MH  - Amino Acid Sequence
MH  - Carcinoma, Renal Cell/immunology
MH  - Cell Line, Tumor
MH  - Cells/immunology
MH  - Flow Cytometry
MH  - Histocompatibility Antigens Class I/chemistry/*isolation & purification
MH  - Humans
MH  - Kidney Neoplasms/immunology
MH  - Major Histocompatibility Complex
MH  - Neoplasm Proteins/chemistry/*isolation & purification
MH  - Peptide Fragments/chemistry/isolation & purification
MH  - Sensitivity and Specificity
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
EDAT- 2005/08/23 09:00
MHDA- 2006/03/29 09:00
CRDT- 2005/08/23 09:00
PHST- 2005/08/23 09:00 [pubmed]
PHST- 2006/03/29 09:00 [medline]
PHST- 2005/08/23 09:00 [entrez]
AID - S1535-9476(20)30025-6 [pii]
AID - 10.1074/mcp.M500076-MCP200 [doi]
PST - ppublish
SO  - Mol Cell Proteomics. 2005 Dec;4(12):1888-97. doi: 10.1074/mcp.M500076-MCP200. 
      Epub 2005 Aug 19.