title: Analysis of formalin-fixed, paraffin-embedded (FFPE) tissue via proteomic techniques and misconceptions of antigen retrieval.
abstract: Since emerging in the late 19(th) century, formaldehyde fixation has become a standard method for preservation of tissues from clinical samples. The advantage of formaldehyde fixation is that fixed tissues can be stored at room temperature for decades without concern for degradation. This has led to the generation of huge tissue banks containing thousands of clinically significant samples. Here we review techniques for proteomic analysis of formalin-fixed, paraffin-embedded (FFPE) tissue samples with a specific focus on the methods used to extract and break formaldehyde crosslinks. We also discuss an error-of-interpretation associated with the technique known as "antigen retrieval." We have discovered that this term has been mistakenly applied to two disparate molecular techniques; therefore, we argue that a terminology change is needed to ensure accurate reporting of experimental results. Finally, we suggest that more investigation is required to fully understand the process of formaldehyde fixation and its subsequent reversal.
authors.affiliation: Proteomics Core Facility, University of Technology Sydney, Cnr Harris and Thomas St, Ultimo, NSW, Australia.
authors.forename: Matthew B
authors.initials: MB
authors.lastname: O'Rourke
authors.affiliation: Proteomics Core Facility, University of Technology Sydney, Cnr Harris and Thomas St, Ultimo, NSW, Australia.
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Padula
mesh_terms: D000941:Antigens / Q000737:chemistry / Q000302:isolation & purification*; D005404:Fixatives / Q000737:chemistry*; D005557:Formaldehyde / Q000737:chemistry*; D020869:Gene Expression Profiling; D016612:Paraffin Embedding; D040901:Proteomics / Q000379:methods*; D016707:Tissue Fixation
chemical_list: D000941:Antigens; D005404:Fixatives; D005557:Formaldehyde
keywords: antigen retrieval; formaldehyde fixation; mass spectrometry; proteomics