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Gruver, Aaron M; Liu, Ling; Vaillancourt, Peter; Yan, Sau‐Chi B; Cook, Joel D; Roseberry Baker, Jessica A; Felke, Erin M; Lacy, Megan E; Marchal, Christophe C; Szpurka, Hadrian; Holzer, Timothy R.; Rhoads, Emily K; Zeng, Wei; Wortinger, Mark A; Lu, Jirong; Chow, Chi‐kin; Denning, Irene J; Beuerlein, Gregory; Davies, Julian; Hanson, Jeff C; Credille, Kelly M; Wijayawardana, Sameera R; Schade, Andrew E

Immunohistochemical application of a highly sensitive and specific murine monoclonal antibody recognising the extracellular domain of the human hepatocyte growth factor receptor (MET)

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  • Medientyp: E-Artikel
  • Titel: Immunohistochemical application of a highly sensitive and specific murine monoclonal antibody recognising the extracellular domain of the human hepatocyte growth factor receptor (MET)
  • Beteiligte: Gruver, Aaron M; Liu, Ling; Vaillancourt, Peter; Yan, Sau‐Chi B; Cook, Joel D; Roseberry Baker, Jessica A; Felke, Erin M; Lacy, Megan E; Marchal, Christophe C; Szpurka, Hadrian; Holzer, Timothy R.; Rhoads, Emily K; Zeng, Wei; Wortinger, Mark A; Lu, Jirong; Chow, Chi‐kin; Denning, Irene J; Beuerlein, Gregory; Davies, Julian; Hanson, Jeff C; Credille, Kelly M; Wijayawardana, Sameera R; Schade, Andrew E
  • Erschienen: Wiley, 2014
  • Erschienen in: Histopathology
  • Sprache: Englisch
  • DOI: 10.1111/his.12510
  • ISSN: 0309-0167; 1365-2559
  • Schlagwörter: General Medicine ; Histology ; Pathology and Forensic Medicine
  • Entstehung:
  • Anmerkungen:
  • Beschreibung: <jats:sec><jats:title>Aims</jats:title><jats:p>Development of novel targeted therapies directed against hepatocyte growth factor (<jats:styled-content style="fixed-case">HGF</jats:styled-content>) or its receptor (<jats:styled-content style="fixed-case">MET</jats:styled-content>) necessitates the availability of quality diagnostics to facilitate their safe and effective use. Limitations of some commercially available anti‐<jats:styled-content style="fixed-case">MET</jats:styled-content> antibodies have prompted development of the highly sensitive and specific clone A2H2‐3. Here we report its analytical properties when applied by an automated immunohistochemistry method.</jats:p></jats:sec><jats:sec><jats:title>Methods and results</jats:title><jats:p>Excellent antibody specificity was demonstrated by immunoblot, <jats:styled-content style="fixed-case">ELISA</jats:styled-content>, and <jats:styled-content style="fixed-case">IHC</jats:styled-content> evaluation of characterised cell lines including <jats:styled-content style="fixed-case">NIH</jats:styled-content>3T3 overexpressing the related kinase <jats:italic><jats:styled-content style="fixed-case">MST</jats:styled-content>1R</jats:italic> (<jats:styled-content style="fixed-case">RON</jats:styled-content>). Sensitivity was confirmed by measurements of <jats:italic><jats:styled-content style="fixed-case">MET</jats:styled-content></jats:italic> in cell lines or characterised tissues. <jats:styled-content style="fixed-case">IHC</jats:styled-content> correlated well with <jats:styled-content style="fixed-case">FISH</jats:styled-content> and quantitative <jats:styled-content style="fixed-case">RT</jats:styled-content>‐<jats:styled-content style="fixed-case">PCR</jats:styled-content> assessments of <jats:italic><jats:styled-content style="fixed-case">MET</jats:styled-content></jats:italic> (<jats:italic>P </jats:italic>&lt;<jats:italic> </jats:italic>0.001). Good total agreement (89%) was observed with the anti‐<jats:styled-content style="fixed-case">MET</jats:styled-content> antibody clone <jats:styled-content style="fixed-case">SP</jats:styled-content>44 using whole‐tissue sections, but poor positive agreement (21–47%) was seen in tissue microarray cores. Multiple lots displayed appropriate reproducibility (<jats:italic>R</jats:italic><jats:sup>2</jats:sup> &gt; 0.9). Prevalence of <jats:styled-content style="fixed-case">MET</jats:styled-content> positivity by <jats:styled-content style="fixed-case">IHC</jats:styled-content> was higher in non‐squamous cell <jats:styled-content style="fixed-case">NSCLC</jats:styled-content>,<jats:italic> <jats:styled-content style="fixed-case">MET</jats:styled-content></jats:italic> or <jats:italic><jats:styled-content style="fixed-case">EGFR</jats:styled-content></jats:italic> amplified cases, and in tumours harbouring abnormalities in <jats:italic><jats:styled-content style="fixed-case">EGFR</jats:styled-content></jats:italic> exon 19 or 21.</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>The anti‐<jats:styled-content style="fixed-case">MET</jats:styled-content> antibody clone A2H2‐3 displays excellent specificity and sensitivity. These properties make it suitable for clinical trial investigations and development as a potential companion diagnostic.</jats:p></jats:sec>