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Immunohistochemical application of a highly sensitive and specific murine monoclonal antibody recognising the extracellular domain of the human hepatocyte growth factor receptor ( MET )
Author(s) -
Gruver Aaron M,
Liu Ling,
Vaillancourt Peter,
Yan SauChi 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 Chikin,
Denning Irene J,
Beuerlein Gregory,
Davies Julian,
Hanson Jeff C,
Credille Kelly M,
Wijayawardana Sameera R,
Schade Andrew E
Publication year - 2014
Publication title -
histopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.626
H-Index - 124
eISSN - 1365-2559
pISSN - 0309-0167
DOI - 10.1111/his.12510
Subject(s) - immunohistochemistry , clone (java method) , hepatocyte growth factor receptor , antibody , monoclonal antibody , biology , hepatocyte growth factor , tissue microarray , monoclonal , epidermal growth factor receptor , pathology , cancer research , microbiology and biotechnology , receptor , medicine , c met , immunology , gene , biochemistry
Aims Development of novel targeted therapies directed against hepatocyte growth factor ( HGF ) or its receptor ( MET ) necessitates the availability of quality diagnostics to facilitate their safe and effective use. Limitations of some commercially available anti‐ MET 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. Methods and results Excellent antibody specificity was demonstrated by immunoblot, ELISA , and IHC evaluation of characterised cell lines including NIH 3T3 overexpressing the related kinase MST 1R ( RON ). Sensitivity was confirmed by measurements of MET in cell lines or characterised tissues. IHC correlated well with FISH and quantitative RT ‐ PCR assessments of MET ( P < 0.001). Good total agreement (89%) was observed with the anti‐ MET antibody clone SP 44 using whole‐tissue sections, but poor positive agreement (21–47%) was seen in tissue microarray cores. Multiple lots displayed appropriate reproducibility ( R 2 > 0.9). Prevalence of MET positivity by IHC was higher in non‐squamous cell NSCLC , MET or EGFR amplified cases, and in tumours harbouring abnormalities in EGFR exon 19 or 21. Conclusions The anti‐ MET 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.