Revealing the SARS-CoV-2 Spike Protein and Specific Antibody Immune Complex Formation Mechanism for Precise Evaluation of Antibody Affinity.

The profound understanding and detailed evaluation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (SCoV2-S) protein and specific antibody interaction mechanism is of high importance in the development of immunosensors for COVID-19. In the present work, we studied a model system of immobilized SCoV2-S protein and specific monoclonal antibodies by molecular dynamics of immune complex formation in real time. We simultaneously applied spectroscopic ellipsometry and quartz crystal microbalance with dissipation to reveal the features and steps of the immune complex formation. We showed direct experimental evidence based on acoustic and optical measurements that the immune complex between covalently immobilized SCoV2-S and specific monoclonal antibodies is formed in two stages. Based on these findings it was demonstrated that applying a two-step binding mathematical model for kinetics analysis leads to a more precise determination of interaction rate constants than that determined by the 1:1 Langmuir binding model. Our investigation showed that the equilibrium dissociation constants (KD) determined by a two-step binding model and the 1:1 Langmuir model could differ significantly. The reported findings can facilitate a deeper understanding of antigen-antibody immune complex formation steps and can open a new way for the evaluation of antibody affinity towards corresponding antigens.

Biosensor Based on Graphene Directly Grown by MW-PECVD for Detection of COVID-19 Spike (S) Protein and Its Entry Receptor ACE2.

Biosensors based on graphene field-effect transistors (G-FET) for detecting COVID-19 spike S protein and its receptor ACE2 were reported. The graphene, directly synthesized on SiO2/Si substrate by microwave plasma-enhanced chemical vapor deposition (MW-PECVD), was used for FET biosensor fabrication. The commercial graphene, CVD-grown on a copper substrate and subsequently transferred onto a glass substrate, was applied for comparison purposes. The graphene structure and surface morphology were studied by Raman scattering spectroscopy and atomic force microscope. Graphene surfaces were functionalized by an aromatic molecule PBASE (1-pyrenebutanoic acid succinimidyl ester), and subsequent immobilization of the receptor angiotensin-converting enzyme 2 (ACE2) was performed. A microfluidic system was developed, and transfer curves of liquid-gated FET were measured after each graphene surface modification procedure to investigate ACE2 immobilization by varying its concentration and subsequent spike S protein detection. The directly synthesized graphene FET sensitivity to the receptor ACE2, evaluated in terms of the Dirac voltage shift, exceeded the sensitivity of the transferred commercial graphene-based FET. The concentration of the spike S protein was detected in the range of 10 ag/mL up to 10 µg/mL by using a developed microfluidic system and measuring the transfer characteristics of the liquid-gated G-FETs. It was found that the shift of the Dirac voltage depends on the spike S concentration and was 27 mV with saturation at 10 pg/mL for directly synthesized G-FET biosensor, while for transferred G-FET, the maximal shift of 70 mV was obtained at 10 µg/mL with a tendency of saturation at 10 ng/mL. The detection limit as low as 10 ag/mL was achieved for both G-FETs. The sensitivity of the biosensors at spike S concentration of 10 pg/mL measured as relative current change at a constant gate voltage corresponding to the highest transconductance of the G-FETs was found at 5.6% and 8.8% for directly synthesized and transferred graphene biosensors, respectively. Thus, MW-PECVD-synthesized graphene-based biosensor demonstrating high sensitivity and low detection limit has excellent potential for applications in COVID-19 diagnostics.

Measurement System for Short-Pulsed Magnetic Fields.

A measurement system based on the colossal magnetoresistance CMR-B-scalar sensor was developed for the measurement of short-duration high-amplitude magnetic fields. The system consists of a magnetic field sensor made from thin nanostructured manganite film with minimized memory effect, and a magnetic field recording module. The memory effect of the La1-xSrx(Mn1-yCoy)zO3 manganite films doped with different amounts of Co and Mn was investigated by measuring the magnetoresistance (MR) and resistance relaxation in pulsed magnetic fields up to 20 T in the temperature range of 80-365 K. It was found that for low-temperature applications, films doped with Co (LSMCO) are preferable due to the minimized magnetic memory effect at these temperatures, compared with LSMO films without Co. For applications at temperatures higher than room temperature, nanostructured manganite LSMO films with increased Mn content above the stoichiometric level have to be used. These films do not exhibit magnetic memory effects and have higher MR values. To avoid parasitic signal due to electromotive forces appearing in the transmission line of the sensor during measurement of short-pulsed magnetic fields, a bipolar-pulsed voltage supply for the sensor was used. For signal recording, a measurement module consisting of a pulsed voltage generator with a frequency up to 12.5 MHz, a 16-bit ADC with a sampling rate of 25 MHz, and a microprocessor was proposed. The circuit of the measurement module was shielded against low- and high-frequency electromagnetic noise, and the recorded signal was transmitted to a personal computer using a fiber optic link. The system was tested using magnetic field generators, generating magnetic fields with pulse durations ranging from 3 to 20 µs. The developed magnetic field measurement system can be used for the measurement of high-pulsed magnetic fields with pulse durations in the order of microseconds in different fields of science and industry.

The Application of a CMR-B-Scalar Sensor for the Investigation of the Electromagnetic Acceleration of Type II Superconductors.

In this paper, we investigated the behavior of a type II superconducting armature when accelerated by a pulsed magnetic field generated by a single-stage pancake coil. While conducting this investigation, we performed a numerical finite element simulation and an experimental study of the magnetic field dynamics at the edge of the pancake coil when the payload was a superconducting disc made from YBa2Cu3O7-x, cooled down to 77 K. The magnetic field measurements were performed using a CMR-B-scalar sensor, which was able to measure the absolute magnitude of the magnetic field and was specifically manufactured in order to increase the sensor's sensitivity up to 500 mT. It was obtained that type II superconducting armatures can outperform normal metals when the launch conditions are tailored to their electromagnetic properties.