Abstract
Hemoglobin is an iron carrying protein in erythrocytes and also an essential element to transfer oxygen from the lungs to the tissues. Abnormalities in hemoglobin concentration are closely correlated with health status and many diseases, including thalassemia, anemia, leukemia, heart disease, and excessive loss of blood. Particularly in resource-constrained settings existing blood analyzers are not readily applicable due to the need for high-level instrumentation and skilled personnel, thereby inexpensive, easy-to-use, and reliable detection methods are needed. Herein, a molecular fingerprints of hemoglobin on a nanofilm chip was obtained for real-time, sensitive, and selective hemoglobin detection using a surface plasmon resonance system. Briefly, through the photopolymerization technique, a template (hemoglobin) was imprinted on a monomeric (acrylamide) nanofilm on-chip using a cross-linker (methylenebisacrylamide) and an initiator-activator pair (ammonium persulfate-tetramethylethylenediamine). The molecularly imprinted nanofilm on-chip was characterized by atomic force microscopy and ellipsometry, followed by benchmarking detection performance of hemoglobin concentrations from 0.0005 mg mL−1 to 1.0 mg mL−1. Theoretical calculations and real-time detection implied that the molecularly imprinted nanofilm on-chip was able to detect as little as 0.00035 mg mL−1 of hemoglobin. In addition, the experimental results of hemoglobin detection on the chip well-fitted with the Langmuir adsorption isotherm model with high correlation coefficient (0.99) and association and dissociation coefficients (39.1 mL mg−1 and 0.03 mg mL−1) suggesting a monolayer binding characteristic. Assessments on selectivity, reusability and storage stability indicated that the presented chip is an alternative approach to current hemoglobin-targeted assays in low-resource regions, as well as antibody-based detection procedures in the field. In the future, this molecularly imprinted nanofilm on-chip can easily be integrated with portable plasmonic detectors, improving its access to these regions, as well as it can be tailored to detect other proteins and biomarkers.
Highlights
Proteins display multiple functional and structural features in cellular machinery
The molecularly imprinted nanofilm on-chip was characterized by atomic force microscopy and ellipsometry, followed by benchmarking detection performance of hemoglobin concentrations from 0.0005 mg mL−1 to 1.0 mg mL−1
The pre-complex was prepared with the ratio 1 μmol:4 mmol of hemoglobin and acrylamide and the absorbance intensity increment ended at 1this ratio.mmol
Summary
Proteins display multiple functional and structural features in cellular machinery. They are essential elements as indicators and predictors of diseases through their inherent structural properties, concentration, and 3-D orientation [1]. Such a compound, hemoglobin, a tetrameric protein in red blood cells, consists of two dimer subunits with an iron-carrying protein that transports oxygen and carbon dioxide throughout the body and maintains acid-base balance in the blood. There are many laboratory-based analytical tools to determine hemoglobin levels, considerable limitations including expensive assays, multi-step procedures, long assay times, the need for skilled personnel, poor test stability and specificity still limit their implementations in resource-constrained. Even though many efforts are directed at developing sensitive detection strategies, especially affinity-based methods, transportation, refrigeration, and storage are main issues in these regions, thereby easy-to-use, reliable, sensitive, durable, specific and long-term stable detection assays are urgently needed [5]
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