1O - Detection of ALK rearranged non-small cell lung carcinomas by immunohistochemistry: comparison of different antibodies (D5F3 and 5A4), detection sys...

Date 27 March 2014
Event ELCC 2014
Session Proffered Papers 2 - Tumour biology and translational research
Topics Lung and other Thoracic Tumours
Pathology/Molecular Biology
Presenter Spasenija Savic
Citation Journal of Thoracic Oncology (2014) 9 (Supplement 9): S7-S52. 10.1097/JTO.0000000000000131
Authors S. Savic1, J. Diebold2, A. Zimmermann3, W. Jochum4, B. Baschiera1, R. Epper1, S. Grieshaber1, L. Tornillo1, K. Kerr5, L. Bubendorf1
  • 1Pathology, University Hospital Basel, 4056 - Basel/CH
  • 2Institute Of Pathology, Cantonal Hospital Lucerne, Luzern/CH
  • 3Institute Of Surgical Pathology, University Hospital Zurich, Zürich/CH
  • 4Institute Of Pathology, Cantonal Hospital St.Gallen, St. Gallen/CH
  • 5Department Of Pathology, Aberdeen Royal Infirmary Anchor Unit, Aberdeen/UK


Immunohistochemistry (IHC) has become a promising method for prescreening predictive ALK-rearrangements in non-small cell lung carcinomas (NSCLC). Various ALK antibodies, enhanced detection systems and automated immunostainers are available. We therefore aimed to compare the performance of: 1. the D5F3 (Cell Signaling, Ventana) and the 5A4 (Novocastra, Leica) monoclonal antibodies (mAb) and 2. the 5A4 mAb on two different automated immunostainers (BenchMark, Ventana and BOND-MAX, Leica).

Methods: Sixty-five formalin-fixed and paraffin-embedded non-squamous NSCLC with available ALK FISH results were retrospectively analysed. The specimens were enriched for ALK FISH-positive NSCLC (24/65, 36.9%). IHC was performed with the OptiView Amplification Kit (Ventana) on BenchMark using the D5F3 and the 5A4 mAb, respectively. Additionally IHC with the 5A4 mAb and the Bond Polymer Refine Detection Kit (Leica) on Bond-MAX was performed. For D5F3 any strong IHC-staining (3+) was regarded as a positive result, as suggested by the manufacturer. For 5A4 any IHC-staining (1+, 2+, 3+) was regarded as a positive result. The IHC results of all three protocols were compared to the FISH results. In case of a discrepancy the IHC-stained slide and the FISH signals were reviewed.