Fast and efficient isolation of human EGFR-positive cells using EasySep&[trade]

• Published in: The Journal of immunology (1950) Vol. 210; no. 1_Supplement; pp. 62 - 62.05
• Main Authors: Poon, Grace FT, Liang, Alice, Chehal, Manreet, Eaves, Allen C, Louis, Sharon A, Antignano, Frann
• Format: Journal Article
• Language: English
• Published: 01-05-2023
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.210.Supp.62.05

Abstract
Epidermal growth factor receptor (EGFR) is a cell surface receptor normally expressed on most epithelial cells and some hematopoietic cells. Mutation and overexpression of EGFR is a key driver of many human cancers, including lung, breast, as well as colorectal cancers. Circulating tumour cells (CTCs) can also express EGFR, where enrichment of EGFR+ CTCs can provide promising liquid biopsy and prognostic values. EGFR-positive cells can be difficult to isolate because they are present in many tissue types at a wide range of frequencies. We have developed a simple method to isolate human EGFR-positive cells to address this challenge. A549 human lung adenocarcinoma cells expressing EGFR were spiked into human PBMCs at defined frequencies as a model system. Starting with a single-cell suspension with 10% EGFR+ cells, the EGFR-expressing cells were labeled with an antibody complex that linked the EGFR-expressing cells to magnetic particles, then separated using an EasySep™ magnet. Using this method, EGFR+ cells were enriched from 9.9 ± 3.3% to 95.0 ± 2.8% (mean ± SD; n = 14). Protocols have been optimized for different sample sizes and different EGFR+ cell starting frequencies. EasySep™-isolated EGFR+ lung adenocarcinoma cells were viable and proliferative, with fold expansion and Ki67 expression comparable to untreated cells. EasySep™ Human EGFR Positive Selection Kit enables simple and easy isolation of EGFR-positive cells in 15 minutes, facilitating EGFR and cancer research.