title: Label-free capture of breast cancer cells spiked in buffy coats using carbon nanotube antibody micro-arrays.
abstract: We demonstrate the rapid and label-free capture of breast cancer cells spiked in buffy coats using nanotube-antibody micro-arrays. Single wall carbon nanotube arrays were manufactured using photo-lithography, metal deposition, and etching techniques. Anti-epithelial cell adhesion molecule (EpCAM) antibodies were functionalized to the surface of the nanotube devices using 1-pyrene-butanoic acid succinimidyl ester functionalization method. Following functionalization, plain buffy coat and MCF7 cell spiked buffy coats were adsorbed on to the nanotube device and electrical signatures were recorded for differences in interaction between samples. A statistical classifier for the 'liquid biopsy' was developed to create a predictive model based on dynamic time warping to classify device electrical signals that corresponded to plain (control) or spiked buffy coats (case). In training test, the device electrical signals originating from buffy versus spiked buffy samples were classified with ∼100% sensitivity, ∼91% specificity and ∼96% accuracy. In the blinded test, the signals were classified with ∼91% sensitivity, ∼82% specificity and ∼86% accuracy. A heatmap was generated to visually capture the relationship between electrical signatures and the sample condition. Confocal microscopic analysis of devices that were classified as spiked buffy coats based on their electrical signatures confirmed the presence of cancer cells, their attachment to the device and overexpression of EpCAM receptors. The cell numbers were counted to be ∼1-17 cells per 5 μl per device suggesting single cell sensitivity in spiked buffy coats that is scalable to higher volumes using the micro-arrays.
authors.affiliation: Department of Mechanical Engineering, Small Systems Laboratory, Worcester Polytechnic Institute, Worcester, MA 01532, USA.
authors.forename: Farhad
authors.initials: F
authors.lastname: Khosravi
authors.forename: Patrick
authors.initials: P
authors.lastname: Trainor
authors.forename: Shesh N
authors.initials: SN
authors.lastname: Rai
authors.forename: Goetz
authors.initials: G
authors.lastname: Kloecker
authors.forename: Eric
authors.initials: E
authors.lastname: Wickstrom
authors.forename: Balaji
authors.initials: B
authors.lastname: Panchapakesan
mesh_terms: D000906:Antibodies / Q000378:metabolism*; D001943:Breast Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D005260:Female; D006801:Humans; D061986:MCF-7 Cells; D037742:Nanotubes, Carbon / Q000737:chemistry; D040081:Protein Array Analysis / Q000379:methods*; D013194:Staining and Labeling
chemical_list: D000906:Antibodies; D037742:Nanotubes, Carbon