title: Proteome analysis of microdissected formalin-fixed and paraffin-embedded tissue specimens.
abstract: Targeted proteomics research, based on the enrichment of disease-relevant proteins from isolated cell populations selected from high-quality tissue specimens, offers great potential for the identification of diagnostic, prognostic, and predictive biological markers for use in the clinical setting and during preclinical testing and clinical trials, as well as for the discovery and validation of new protein drug targets. Formalin-fixed and paraffin-embedded (FFPE) tissue collections, with attached clinical and outcome information, are invaluable resources for conducting retrospective protein biomarker investigations and performing translational studies of cancer and other diseases. Combined capillary isoelectric focusing/nano-reversed-phase liquid chromatography separations equipped with nano-electrospray ionization-tandem mass spectrometry are employed for the studies of proteins extracted from microdissected FFPE glioblastoma tissues using a heat-induced antigen retrieval (AR) technique. A total of 14,478 distinct peptides are identified, leading to the identification of 2733 non-redundant SwissProt protein entries. Eighty-three percent of identified FFPE tissue proteins overlap with those obtained from the pellet fraction of fresh-frozen tissue of the same patient. This large degree of protein overlapping is attributed to the application of detergent-based protein extraction in both the cell pellet preparation protocol and the AR technique.
authors.affiliation: Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland, USA.
authors.forename: Tong
authors.initials: T
authors.lastname: Guo
authors.forename: Weijie
authors.initials: W
authors.lastname: Wang
authors.forename: Paul A
authors.initials: PA
authors.lastname: Rudnick
authors.forename: Tao
authors.initials: T
authors.lastname: Song
authors.forename: Jie
authors.initials: J
authors.lastname: Li
authors.forename: Zhengping
authors.initials: Z
authors.lastname: Zhuang
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Weil
authors.forename: Don L
authors.initials: DL
authors.lastname: DeVoe
authors.forename: Cheng S
authors.initials: CS
authors.lastname: Lee
authors.forename: Brian M
authors.initials: BM
authors.lastname: Balgley
mesh_terms: D000595:Amino Acid Sequence; D001932:Brain Neoplasms / Q000378:metabolism; D002853:Chromatography, Liquid; D005189:False Positive Reactions; D005404:Fixatives; D005557:Formaldehyde; D005909:Glioblastoma / Q000378:metabolism; D006801:Humans; D007525:Isoelectric Focusing; D042282:Microdissection; D008969:Molecular Sequence Data; D016612:Paraffin Embedding; D010455:Peptides / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D005404:Fixatives; D010455:Peptides; D020543:Proteome; D005557:Formaldehyde