title: Anti-EpCAM modified LC-SPDP monolayer on gold microelectrode based electrochemical biosensor for MCF-7 cells detection.
abstract: A succinimidyl 6-(3-[2-pyridyldithio]-propionamido) hexanoate (LC-SPDP) self-assembled monolayer (SAM) prepared onto a 500 μm (diameter) gold microelectrode (Au) surface has been utilized for covalent immobilization of anti-EpCAM antibody. Amino group on anti-EpCAM antibody was covalently bound with succinimidyl group on SAM via amide bond and unreacted active groups of LC-SPDP were blocked using 1% ethanol amine (EA). These anti-EpCAM/LC-SPDP/Au electrodes were characterized using cyclic voltammetric (CV) and fluorescence techniques, respectively. The anti-EpCAM/LC-SPDP/Au electrodes were exposed to solutions with different MCF-7 cell concentrations and CV technique was used to determine the cell concentration. Further, CV studies on blank 500 and 50 μm (diameter) gold microelectrodes were used to identify cell via molecular profiling using ferrocene amidopentyl carboxylic acid based redox tagging and magnetic beads based enhancement. CV results confirm that the anti-EpCAM/LC-SPDP/Au based biosensor could detect MCF-7 cells in the range of 1×10(5)-1×10(8) with correlation coefficient of 0.999 and detection limit of 1×10(5) cells ml(-1) i.e. 100 cells in solution used for incubation (1 μl). Molecular profiling studies suggest that smaller size microelectrode (50 μm; diameter) with magnetic beads based enhancement can be employed to identify cell type. This work establishes the feasibility of using microelectrode based platform for breast cancer specific MCF-7 cell concentration estimation and their molecular profiling.
authors.affiliation: Bioelectronics Program, Institute of Microelectronics, A*STAR (Agency for Science, Technology and Research), 11 Science Park Road, Singapore Science Park II, Singapore 117685, Singapore. sunilarya333@gmail.com
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Arya
authors.forename: Karen Yanping
authors.initials: KY
authors.lastname: Wang
authors.forename: Chee Chung
authors.initials: CC
authors.lastname: Wong
authors.forename: Abdur Rub Abdur
authors.initials: AR
authors.lastname: Rahman
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000276:immunology*; D015374:Biosensing Techniques / Q000295:instrumentation*; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000276:immunology*; D045744:Cell Line, Tumor; D003217:Conductometry / Q000295:instrumentation*; D003581:Cytodiagnosis / Q000295:instrumentation*; D000071858:Epithelial Cell Adhesion Molecule; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D008839:Microelectrodes; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D006046:Gold