A novel, high-throughput workflow for discovery and identification of serum carrier protein-bound peptide biomarker candidates in ovarian cancer samples

Mary F. Lopez*, Alvydas Mikulskis, Scott Kuzdzal, Eva Golenko, Emanuel F. Petricoin, Lance A. Liotta, Wayne F. Patton, Gordon R. Whiteley, Kevin Rosenblatt, Prem Gurnani, Animesh Nandi, Samuel Neill, Stuart Cullen, Martin O'Gorman, David Sarracino, Christopher Lynch, Andrew Johnson, William Mckenzie, David Fishman

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

106 Scopus citations

Abstract

Background: Most cases of ovarian cancer are detected at later stages when the 5-year survival is ∼15%, but 5-year survival approaches 90% when the cancer is detected early (stage I). To use mass spectrometry (MS) of serum proteins for early detection, a seamless workflow is needed that provides an opportunity for rapid profiling along with direct identification of the underpinning ions. Methods: We used carrier protein-bound affinity enrichment of serum samples directly coupled with MALDI orthagonal TOF MS profiling to rapidly search for potential ion signatures that contained discriminatory power. These ions were subsequently directly subjected to tandem MS for sequence identification. Results: We discovered several biomarker panels that enabled differentiation of stage I ovarian cancer from unaffected (age-matched) patients with no evidence of ovarian cancer, with positive results in >93% of samples from patients with disease-negative results and in 97% of disease-free controls. The carrier protein-based approach identified additional protein fragments, many from low-abundance proteins or proteins not previously seen in serum. Conclusions: This workflow system using a highly reproducible, high-resolution MALDI-TOF platform enables rapid enrichment and profiling of large numbers of clinical samples for discovery of ion signatures and integration of direct sequencing and identification of the ions without need for additional offline, time-consuming purification strategies.

Original languageEnglish
Pages (from-to)1067-1074
Number of pages8
JournalClinical Chemistry
Volume53
Issue number6
DOIs
StatePublished - Jun 2007
Externally publishedYes

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