Photoluminescence-based biosensor for the detection of antibodies against SARS-CoV-2 virus proteins by ZnO tetrapod structure integrated within microfluidic system.

This paper reports on development of an optical biosensor for the detection of antibodies against SARS-CoV-2 virus proteins in blood serum. ZnO nanotetrapods with high surface area and stable room temperature photoluminescence (PL) were selected as transducers. Structure and optical properties of the ZnO tetrapods have been studied by XRD, SEM and Raman spectroscopy. Crystallinity, dimensions and emission peaks of the ZnO tetrapods were determined. The ZnO tetrapods were fixed on glass chip. Silanization of ZnO tetrapods surface resulted in forming of functional surface groups suitable for the immobilization of bioselective layer. Two types of recombinant proteins (rS and rN) have been used to form bioselective layer on the surface of the ZnO tetrapods. Flow through microfluidic system, integrated with optical system, has been used for the determination of antibodies against SARS-CoV-2 virus proteins present in blood samples. The SARS-CoV-2 probes, prepared in PBS solution, have been injected into the measurement chamber with a constant pumping speed. Steady-state photoluminescence spectra and photoluminescence kinetics have been studied before and after injection of the probes. The biosensor signal has been tested to anti-SARS-CoV-2 antibodies in the range of 0.001 nM-1 nM. Control measurements have been performed with blood serum of healthy person. ZnO-SARS-CoV-2-rS and ZnO-SARS-CoV-2-rN biosensors showed high stability and sensitivity to anti-SARS-CoV-2 antibodies in the range of 0.025-0.5 nM (LOD 0.01 nM) and 0.3-1 nM (LOD 0.3 nM), respectively. Gibbs free energy of interaction between ZnO/SARS-CoV-2-rS and ZnO/SARS-CoV-2-rN bioselective layers with anti-SARS-CoV-2 antibodies showed -35.5 and -21.4 kJ/mol, respectively. Average detection time of biosensor integrated within microfluidic system was 15-20 min. The detection time and pumping speed (50 µL/min) were optimized to make detection faster. The developed system and ZnO-SARS-CoV-2-rS nanostructures have good potential for detection of anti-SARS-CoV-2 antibodies from patient's probes.

Molecular imprinting technology for biomedical applications.

Molecularly imprinted polymers (MIPs), a type of biomimetic material, have attracted considerable interest owing to their cost-effectiveness, good physiochemical stability, favourable specificity and selectivity for target analytes, and widely used for various biological applications. It was demonstrated that MIPs with significant selectivity towards protein-based targets could be applied in medicine, diagnostics, proteomics, environmental analysis, sensors, various in vivo and/or in vitro applications, drug delivery systems, etc. This review provides an overview of MIPs dedicated to biomedical applications and insights into perspectives on the application of MIPs in newly emerging areas of biotechnology. Many different protocols applied for the synthesis of MIPs are overviewed in this review. The templates used for molecular imprinting vary from the minor glycosylated glycan-based structures, amino acids, and proteins to whole bacteria, which are also overviewed in this review. Economic, environmental, rapid preparation, stability, and reproducibility have been highlighted as significant advantages of MIPs. Particularly, some specialized MIPs, in addition to molecular recognition properties, can have high catalytic activity, which in some cases could be compared with other bio-catalytic systems. Therefore, such MIPs belong to the class of so-called 'artificial enzymes'. The discussion provided in this manuscript furnishes a comparative analysis of different approaches developed, underlining their relative advantages and disadvantages highlighting trends and possible future directions of MIP technology.