An ultra-sensitive SARS-CoV-2 antigen optical biosensor based on angiotensin converting enzyme 2 (ACE-2) functionalized magnetic-fluorescent silica nanoparticles.

This work reports on the design and synthesis of an angiotensin-converting enzyme 2 (ACE-2) functionalized magnetic fluorescent silica nanoparticles (Fe-FSNP) as a biosensing platform to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen. Iron oxide (Fe3O4) nanoparticles were synthesized via ultrasonic-assisted coprecipitation and then coated with fluorescent silica nanoparticles (FSNP) through thesol-gelmethod forming the Fe-FSNP samples. Silica obtained from local geothermal powerplant was used in this work and Rhodamine B was chosen as the incorporated fluorescent dye, hence this reports for the first time ACE-2 was immobilized on the natural silica surface. The Fe-FSNP nanoparticle consists of a 18-25 nm magnetic core and a silica shell with a thickness of 30 nm as confirmed from the transmission electron microscopy image. Successful surface functionalization of the Fe-FSNP with ACE-2 as bioreceptor was conducted through hydrosylilation reaction and confirmed through the Fourier transform infrared spectroscopy. The detection of SARS-Cov-2 antigen by Fe-FSNP/ACE2 was measured through the change in its maximum fluorescence intensity at 588 nm where fluorescence- quenching had occurred. The biosensing platform showed a rapid response at 30 min with a linear range of 10-6to 10-2µg ml-1. The magnetic-fluorescent properties of the nanoparticle enables an ultra-sensitive detection of SARS-Cov-2 antigen with the limit of detection as low as 2 fg ml-1.

Optimized antibody immobilization on natural silica-based nanostructures for the selective detection of E. coli.

This study reports for the first time the surface modification of fluorescent nanoparticles derived from geothermal silica precipitate with Escherichia coli (E. coli) antibody. The immobilization of biomolecules on the inorganic surface has been carried out using two different pathways, namely the silanization and hydrosilylation reactions. The former applied (3-aminopropyl)triethoxysilane (APTES) as the crosslinker, while the latter used N-hydroxysuccinimide coupled with N-ethyl-N'-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC/NHS). Fourier transform infrared (FTIR) spectroscopy, field emission scanning electron microscopy with energy dispersive X-ray spectroscopy (FESEM-EDX), and fluorescence spectroscopy were used to confirm the chemical, physical, and optical properties of the surface-modified fluorescent silica nanoparticles (FSNPs). Based on the results of the FTIR, fluorescence spectroscopy and stability tests, the modified FSNPs with EDC/NHS with a ratio of 4 : 1 were proven to provide the optimum results for further conjugation with antibodies, affording the FSNP-Ab2 sample. The FSNP-Ab2 sample was further tested as a nanoplatform for the fluorescence-quenching detection of E. coli, which provided a linear range of 102 to 107 CFU mL-1 for E. coli with a limit of detection (LoD) of 1.6 × 102 CFU mL-1. The selectivity of the biosensor was observed to be excellent for E. coli compared to that for P. aeruginosa and S. typhimurium, with reductions in the maximum fluorescence intensity at 588 nm of 89.22%, 26.23%, and 54.06%, respectively. The inorganic nanostructure-biomolecule conjugation with optimized coupling agents showed promising analytical performance as a selective nanoplatform for detecting E. coli bacteria.