Identification of a naturally presented MAGE-A3 epitope on the surface of HLE cell line by mass spectrometry.
- Author:
Ji-run PENG
1
;
Nan DONG
;
Hu-wei LIU
;
Xi-sheng LENG
Author Information
- Publication Type:Journal Article
- MeSH: Amino Acid Sequence; Antigen Presentation; Antigens, Neoplasm; analysis; isolation & purification; Carcinoma, Hepatocellular; immunology; pathology; Cell Line, Tumor; Chromatography, High Pressure Liquid; Epitopes, T-Lymphocyte; analysis; isolation & purification; HLA-A2 Antigen; immunology; Humans; Liver Neoplasms; immunology; pathology; Mass Spectrometry; Neoplasm Proteins; analysis; isolation & purification
- From: Chinese Journal of Surgery 2007;45(9):595-597
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo identify a naturally presented HLA-A2-restricted epitope of MAGE-A3 antigen, FLWGPRALV (MAGE-A3(271 - 279)), on the surface of a human hepatocellular carcinoma (HCC) cell line HLE.
METHODSSynthetic peptide FLWGPRALV, served as positive control target, was analyzed by HPLC and HPLC-ESI-TOF-MSMS, in order to determine its HPLC elution time, mass-spectrometric characteristics and the lowest detection limitation by the two approaches. 3 x 10(9) HLE cells were collected, peptides naturally presented by major histocompatibility complex (MHC) molecules on the cell surface were isolated by mild acid elution, and concentrated by lyophilization, then the mixtures of peptides were fractioned by HPLC. The ingredient ranged from 2 min before the elution time determined by the synthetic peptide to 2 min after that was collected, concentrated by lyophilization, and analyzed by HPLC-ESI-TOF-MSMS, to identify the existence of the MAGE-A3(271 - 279) peptide.
RESULTSThe HPLC-ESI-TOF-MSMS detection provided an evidence for the existence of a doubly charged ion of (m/z)(2) 529.9, which was further analyzed by collision induced dissociation. The doubly charged ion was ultimately identified as the MAGE-A3(271 - 279) peptide, its amino sequence was FLWGPRALV and its molecular weight was 1058.4 Da.
CONCLUSIONSMAGE-A3(271 - 279) epitope could be naturally presented by HLA-A2 molecules to the surface of HCC cell line and MAGE-A3(271 - 279) peptide may have potential immunotherapeutic value in HCC patients.
