Electrochemical distinction of neuronal and neuroblastoma cells via the phosphorylation of the cellular extracellular membrane.

Anal Biochem

Department of Physical and Environmental Sciences, University of Toronto Scarborough, 1265 Military Trail, Toronto, M1C 1A4, Canada; Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, M5S 3H6, Canada. Electronic address:

Published: May 2022

In this contribution we establish a proof of concept method for monitoring, quantifying and differentiating the extracellular phosphorylation of Human SHSY5Y undifferentiated neuronal cells and neuroblastoma cells by three prominent ectokinases PKA, PKC and Src. Herein it is demonstrated that a combination of different experimental techniques, including fluorescence microscopy, quartz crystal microscopy (QCM) and electrochemistry, can be used to detect extracellular phosphorylation levels of neuronal and neuroblastoma cells. Phosphorylation profiles of the three ectokinases, PKA, PKC and Src, were investigated using fluorescence microscopy and the number of phosphorylation sites per kinase was estimated using QCM. Finally, the phosphorylation of the extracellular membrane was determined using electrochemistry. Our results clearly demonstrate that the extracellular phosphorylation of neuronal cells differs significantly in terms of its phosphorylation profile from diseased neuroblastoma cells and the strength of surface electrochemical techniques in the differentiation process. We reveal that using electrochemistry, the percent compositions of neuronal and neuroblastoma cells can also be identified.

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http://dx.doi.org/10.1016/j.ab.2021.114434DOI Listing

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