A new ligand of CD105 screened out by phage display technology provides a reliable identification of recurrent or metastasizing pleomorphic adenoma from pleomorphic adenoma

Abstract

Objectives: To assess CD105 expression in pleomorphic adenoma (PA), recurrent pleomorphic adenoma (RPA) and metastasizing pleomorphic adenoma (MPA), to identify new epitopes and screen a ligand with high affinity to CD105 by phage display technology, to evaluate the reliability of the new ligand for identifying RPA/MPA from PA.

Methods: Phage display technology was used to screen ligands with high affinity to recombinant human CD105. The ligand with strongest affinity to CD105 was synthesized by FMOC Chemistry according to the sequencing results. The archived formalin fixed paraffin-embedded (FFPE) tissues of 35 PA cases, 12 RPA cases and 2 MPA cases were sliced and immunofluorescent stained. CD105 expression were detected by Confocal laser scanning microscopy (CLSM). The relative fluorescence intensity was calculated with the image processing software Image J. Statistical analyses were performed by the software Graph Pad Prism (Version 7.0a). Using PROC logistic, receiver operating characteristic (ROC) curves, area under ROC curves (AUCs) were generated to assess the sensitivity and specificity of the new ligand for identifying RPA/MPA from PA cases.

Results: A ligand with specialty and high affinity to CD105 i.e. ligand nABPK296 were developed. FITC-labeled ligand nABPK296 confirmed the difference of CD105 expression in RPA/MPA and PA. The AUC of nABPK296 was 0.9418.

Conclusions: CD105 is a promising biomarker for identification of RPA/MPA from PA cases. Ligand nABPK296 provides a promising approach to CD105 detection. This study also validated the reliability of phage display technology in finding new episodes and ligands with high affinity for antigens.

Keywords: Immunofluorescence (IF); Metastasizing pleomorphic adenoma (MPA); Phage display; Pleomorphic adenoma (PA); Recurrent pleomorphic adenoma (RPA).