EGFR Assessment by IHC in Tissue

Methods for EGFR reverse transcription polymerase chain reaction and amplification by fluorescence in situ hybridization^{13 (link)} were previously published. An IHC assay was performed on FFPE tissue to assess EGFR protein expression using the EGFR E30 clone (Agilent, Santa Clara, CA) and the Dako EnVision^TM FLEX detection system on the Dako Link 48 automated staining platform (Agilent). The E30 clone recognizes both wild-type and EGFRvIII forms of EGFR. A trained pathologist evaluated EGFR IHC staining by scoring the percentage of positive cells in neoplastic cells at each intensity, which is categorized into 0 intensity (negative), 1+ intensity (weak), 2+ intensity (moderate), and 3+ intensity (strong).

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Carneiro B.A., Papadopoulos K.P., Strickler J.H., Lassman A.B., Waqar S.N., Chae Y.K., Patel J.D., Shacham-Shmueli E., Kelly K., Khasraw M., Bestvina C.M., Merrell R., Huang K., Atluri H., Ansell P., Li R., Jin J., Anderson M.G., Reilly E.B., Morrison-Thiele G., Patel K., Robinson R.R., Aristide M.R, & Gan H.K. (2022). Phase I study of anti-epidermal growth factor receptor antibody-drug conjugate serclutamab talirine: Safety, pharmacokinetics, and antitumor activity in advanced glioblastoma. Neuro-Oncology Advances, 5(1), vdac183.

Publication 2022

Dako EnVisionTM FLEX detection system

Assay Cells Clone Egfr Egfrviii Fluorescence Neoplastic Pathologist Protein Reverse transcription polymerase chain reaction Tissue Weak

Corresponding Organization : Brown University

Other organizations : South Texas Accelerated Research Therapeutics, Duke Medical Center, Columbia University, NewYork–Presbyterian Hospital, Washington University in St. Louis, Northwestern University, Sheba Medical Center, University of California, Davis, Duke University, University of Chicago, NorthShore University HealthSystem, AbbVie (United States), Austin Health

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Variable analysis

independent variables

EGFR protein expression as measured by IHC using the EGFR E30 clone (Agilent, Santa Clara, CA) and the Dako EnVision™ FLEX detection system on the Dako Link 48 automated staining platform (Agilent)

dependent variables

Percentage of positive cells in neoplastic cells at each intensity (0 intensity (negative), 1+ intensity (weak), 2+ intensity (moderate), and 3+ intensity (strong))

control variables

FFPE tissue samples

controls

The E30 clone recognizes both wild-type and EGFRvIII forms of EGFR, which serves as a positive control for the assay.

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