BACKGROUND: Glycoproteins are often associated with cancer and are important in serum studies, for which glycosylation is a common posttranslational modification. METHODS: We used multilectin affinity chromatography (M-LAC) to isolate glycoproteins from the sera of breast cancer patients and controls. The proteins were identified by HPLC-tandem mass spectrometry (MS/MS) analysis of the corresponding tryptic digests. We used the FuncAssociate Gene Ontology program for association analysis of the identified proteins. Biomarker candidates in these groups were comparatively quantitated by use of peak area measurements, with inclusion of an internal standard. We analyzed data for concordance within the ontology association groups for vector of change with the development of breast cancer. RESULTS: Detection of the known low-concentration biomarker HER-2 (8-24 mug/L) enabled us to establish a dynamic range of 10(6), relative to the amount of albumin, for the depletion step. We then used ELISA to confirm this range. Proteins associated with lipid transport and metabolism, cell growth and maintenance, ion homeostasis, and protease inhibition were found to be differentially regulated in serum from women with breast cancer compared with serum from women without breast cancer. CONCLUSIONS: M-LAC for isolation of the serum glycoproteome, coupled with liquid chromatography-MS/MS and the use of gene ontology associations, can be used to characterize large panels of candidate markers, which can then be evaluated in a particular patient population.
BACKGROUND: Glycoproteins are often associated with cancer and are important in serum studies, for which glycosylation is a common posttranslational modification. METHODS: We used multilectin affinity chromatography (M-LAC) to isolate glycoproteins from the sera of breast cancerpatients and controls. The proteins were identified by HPLC-tandem mass spectrometry (MS/MS) analysis of the corresponding tryptic digests. We used the FuncAssociate Gene Ontology program for association analysis of the identified proteins. Biomarker candidates in these groups were comparatively quantitated by use of peak area measurements, with inclusion of an internal standard. We analyzed data for concordance within the ontology association groups for vector of change with the development of breast cancer. RESULTS: Detection of the known low-concentration biomarker HER-2 (8-24 mug/L) enabled us to establish a dynamic range of 10(6), relative to the amount of albumin, for the depletion step. We then used ELISA to confirm this range. Proteins associated with lipid transport and metabolism, cell growth and maintenance, ion homeostasis, and protease inhibition were found to be differentially regulated in serum from women with breast cancer compared with serum from women without breast cancer. CONCLUSIONS: M-LAC for isolation of the serum glycoproteome, coupled with liquid chromatography-MS/MS and the use of gene ontology associations, can be used to characterize large panels of candidate markers, which can then be evaluated in a particular patient population.
Authors: Zhenxin Lin; Haidi Yin; Andy Lo; Mack T Ruffin; Michelle A Anderson; Diane M Simeone; David M Lubman Journal: Electrophoresis Date: 2013-12-27 Impact factor: 3.535
Authors: Joshua B Owen; Fabio Di Domenico; Rukhsana Sultana; Marzia Perluigi; Chiara Cini; William M Pierce; D Allan Butterfield Journal: J Proteome Res Date: 2009-02 Impact factor: 4.466
Authors: Karen L Abbott; Jae-Min Lim; Lance Wells; Benedict B Benigno; John F McDonald; Michael Pierce Journal: Proteomics Date: 2010-02 Impact factor: 3.984
Authors: Allen D Taylor; William S Hancock; Marina Hincapie; Naoyuki Taniguchi; Samir M Hanash Journal: Genome Med Date: 2009-06-04 Impact factor: 11.117