Liquid Redox Probe-Free Plastic Antibody Development for Malaria Biomarker Recognition

Malaria is a major public health challenge worldwide and requires accurate and efficient diagnostic methods. Traditional diagnostic approaches based on antigen-antibody interactions are associated with ethical and economic concerns. Molecularly imprinted polymers (MIPs) offer a promising alternative by providing a complementary polymer structure capable of selectively binding target molecules. In this study, we developed a liquid, redox-probe-free, MIP-based electrochemical biosensor to detect the Plasmodium falciparum malaria marker histidine-rich protein (HRP2) at the point-of-care (PoC). The imprinting phase consists of the electropolymerization of the monomer methylene blue (MB) in the presence of the target protein HRP2 at the working electrode (WE) of the modified carbon screen printed electrode (C-SPE). Subsequent removal of the protein with proteinase K and oxalic acid yielded the MIP material. The sensor assembly was monitored by cyclic voltammetry (CV), Raman spectroscopy and scanning electron microscopy (SEM). The analytical performance of the biosensor was evaluated by square-wave voltammetry (SWV) using calibration curves in buffer and serum with a detection limit of 0.43 ± 0.026 pg mL^-1. Selectivity studies showed minimal interference, indicating a highly selective assay. Overall, our approach to detect the HRP2 infection marker offers simplicity, cost-effectiveness and reliability. In particular, the absence of a redox solution simplifies detection, as the polymer itself is electroactive and exhibits oxidation and reduction peaks.