Bone, a dynamic tissue with diverse functions in the
human body,
plays vital roles, such as providing structural support and protection
to organs, serving as the site for blood cell formation, and acting
as the primary storage site for calcium. Osteoblasts, bone-forming
cells, express alkaline phosphatase (ALP), which binds to the cell
surface or matrix vesicles and can be released into the bloodstream.
Consequently, ALP can be found in the serum and is commonly used as
a biomarker in clinical studies to assess conditions related to bone
diseases as well as other disorders. This study presents a label-free
approach for detecting ALP using electrochemical impedance spectroscopy
(EIS) using a composite probe consisting of graphene oxide (GO) and
chitosan (Ch)–silk fibroin (SF)–polycaprolactone (PCL)–hydroxyapatite
(HAp) matrix. The GO and composite Ch–SF–PCL–HAp
matrix are sequentially deposited on the glassy carbon electrode and
further immobilized with anti-ALP antibodies. The biosensor probe
is characterized using physical techniques and electrochemical analysis.
The biosensor’s analytical performance is assessed using EIS,
and it shows a limit of detection of 1.74 (±0.26) U/L with a
linear dynamic range of 30–500 U/L, which falls well within
the clinical range of ALP detection. The developed biosensor probe
exhibits high selectivity for ALP (k
sel < 0.04) when tested against interfering molecules in serum. Furthermore,
the system is tested with fetal bovine serum and human serum to detect
ALP in biological samples.