Purification, identification and profiling of serum amyloid A proteins from sera of advanced-stage cancer patients

Jing Li, Zhensheng Xie, Linan Shi, Zhiqiang Zhao, Junjie Hou, Xiulan Chen, Ziyou Cui, Peng Xue, Tanxi Cai, Peng Wu, Sutang Guo, Fuquan Yang

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) is a powerful tool for screening potential biomarkers of various pathological conditions. However, low resolution and mass accuracy of SELDI-TOF-MS remain a major obstacle for determination of biological identities of potential protein biomarkers. We report here a refined workflow that combines ZipTip desalting, acetonitrile precipitation, high-performance liquid chromatography (HPLC) separation and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) analysis for the profiling, purification and identification of the targeted serum proteins found by SELDI-TOF-MS. By using this workflow, we purified ten targeted proteins from the sera of patients with various types of advanced stage (stage III-IV) cancers. These proteins were identified as isoforms of the human serum amyloid protein A (SAA) family with or without truncations at their N-terminals. This was confirmed by Western blot analysis. Different SAA expression patterns were observed by MALDI-TOF-MS profiling. SAA has long been reported as a biomarker for various cancer types such as lung cancer, ovarian cancer, and breast cancer. However, in this study we found increased SAA expression in the sera of advanced-stage cancer patients with different cancer types. Our results suggest that maybe SAA should not be used alone as a biomarker for any specific cancer type.

Original languageEnglish (US)
Pages (from-to)3-9
Number of pages7
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume889-890
DOIs
StatePublished - Mar 15 2012

Keywords

  • MALDI-TOF-MS
  • Purification
  • SAA
  • SELDI-TOF-MS
  • Targeted protein

Fingerprint Dive into the research topics of 'Purification, identification and profiling of serum amyloid A proteins from sera of advanced-stage cancer patients'. Together they form a unique fingerprint.

Cite this