Graphene Oxide Signal Reporter Based Multifunctional
Immunosensing Platform for Amperometric Profiling of Multiple Cytokines
in Serum
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Abstract
Cytokines
are small proteins and form complicated cytokine networks to report
the status of our health. Thus, accurate profiling and sensitive quantification
of multiple cytokines is essential to have a comprehensive and accurate
understanding of the complex physiological and pathological conditions
in the body. In this study, we demonstrated a robust electrochemical
immunosensor for the simultaneous detection of three cytokines IL-6,
IL-1β, and TNF-α. First, graphene oxides (GO) were loaded
with redox probes nile blue (NB), methyl blue (MB), and ferrocene
(Fc), followed by covalent attachment of anti-cytokine antibodies
for IL-6, IL-1β, and TNF-α, respectively, to obtain Ab<sub>2</sub>-GO-NB, Ab<sub>2</sub>-GO-MB, and Ab<sub>2</sub>-GO-Fc, acting
as the signal reporters. The sensing interface was fabricated by attachment
of mixed layers of 4-carboxylic phenyl and 4-aminophenyl phosphorylcholine
(PPC) to glassy carbon surfaces. After that, the capture monoclonal
antibody for IL-6, IL-1β, and TNF-α was modified to the
carboxylic acid terminated sensing interface. And finally a sandwich
assay was developed. The quantitative detection of three cytokines
was achieved by observing the change in electrochemical signal from
signal reporters Ab<sub>2</sub>-GO-NB, Ab<sub>2</sub>-GO-MB, and Ab<sub>2</sub>-GO-Fc. The designed system has been successfully used for
detection of three cytokines (IL-6, IL-1β, and TNF-α)
simultaneously with desirable performance in sensitivity, selectivity,
and stability, and recovery of 93.6%–105.5% was achieved for
determining cytokines spiked in the whole mouse serum