Label-free bead-based metallothionein electrochemical immunosensor
2015
A novel microfluidic label-free bead-based metallothionein
immunosensors was designed. To the surface of superparamagnetic
agarose beads coated with protein A, polyclonal chicken IgY
specifically recognizing metallothionein (MT) were immobilized
via rabbit IgG. The Brdicka reaction was used for
metallothionein detection in a microfluidic printed 3D chip.
Assembled chip consisted of a single copper wire coated with a
thin layer of amalgam as working electrode. Optimization of MT
detection using designed microfluidic chip was performed in
stationary system as well as in the flow arrangement at various
flow rates (0-1800 mikroL/min). In stationary arrangement it is
possible to detect MT concentrations up to 30 ng/mL level, flow
arrangement allows reliable detection of even lower
concentration (12.5 ng/mL). The assembled miniature flow chip
was subsequently tested for the detection of MT elevated levels
(at approx. level 100 mikrog/mL) in real samples of patients
with cancer. The stability of constructed device for
metallothionein detection in flow arrangement was found to be
several days without any maintenance needed.
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