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Targeted Quantitative Mass Spectrometry Analysis of Protein Biomarkers From Previously Stained Single Formalin-Fixed Paraffin-Embedded Tissue Sections.
Ackermann, Bradley L; Morrison, Ryan D; Hill, Salisha; Westfall, Matthew D; Butts, Brent D; Soper, Michael D; Fill, Jeff A; Schade, Andrew E; Liebler, Daniel C; Gruver, Aaron M.
Afiliación
  • Ackermann BL; Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana.
  • Morrison RD; Protypia, Inc, Nashville, Tennessee.
  • Hill S; Protypia, Inc, Nashville, Tennessee.
  • Westfall MD; Protypia, Inc, Nashville, Tennessee.
  • Butts BD; Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana.
  • Soper MD; Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana.
  • Fill JA; Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana.
  • Schade AE; Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana.
  • Liebler DC; Protypia, Inc, Nashville, Tennessee. Electronic address: daniel.liebler@protypia.com.
  • Gruver AM; Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana. Electronic address: gruver_aaron_m@lilly.com.
Lab Invest ; 103(4): 100052, 2023 04.
Article en En | MEDLINE | ID: mdl-36870295
Formalin-fixed, paraffin-embedded tissues represent a majority of all biopsy specimens commonly analyzed by histologic or immunohistochemical staining with adhesive coverslips attached. Mass spectrometry (MS) has recently been used to precisely quantify proteins in samples consisting of multiple unstained formalin-fixed, paraffin-embedded sections. Here, we report an MS method to analyze proteins from a single coverslipped 4-µm section previously stained with hematoxylin and eosin, Masson trichrome, or 3,3'-diaminobenzidine-based immunohistochemical staining. We analyzed serial unstained and stained sections from non-small cell lung cancer specimens for proteins of varying abundance (PD-L1, RB1, CD73, and HLA-DRA). Coverslips were removed by soaking in xylene, and after tryptic digestion, peptides were analyzed by targeted high-resolution liquid chromatography with tandem MS with stable isotope-labeled peptide standards. The low-abundance proteins RB1 and PD-L1 were quantified in 31 and 35 of 50 total sections analyzed, respectively, whereas higher abundance CD73 and HLA-DRA were quantified in 49 and 50 sections, respectively. The inclusion of targeted ß-actin measurement enabled normalization in samples where residual stain interfered with bulk protein quantitation by colorimetric assay. Measurement coefficient of variations for 5 replicate slides (hematoxylin and eosin stained vs unstained) from each block ranged from 3% to 18% for PD-L1, from 1% to 36% for RB1, 3% to 21% for CD73, and 4% to 29% for HLA-DRA. Collectively, these results demonstrate that targeted MS protein quantification can add a valuable data layer to clinical tissue specimens after assessment for standard pathology end points.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Carcinoma de Pulmón de Células no Pequeñas / Neoplasias Pulmonares Tipo de estudio: Guideline Límite: Humans Idioma: En Revista: Lab Invest Año: 2023 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Carcinoma de Pulmón de Células no Pequeñas / Neoplasias Pulmonares Tipo de estudio: Guideline Límite: Humans Idioma: En Revista: Lab Invest Año: 2023 Tipo del documento: Article Pais de publicación: Estados Unidos