Site-Specific Modification of Single Domain Antibodies by Enzyme-Immobilized Magnetic Beads.

Nanobodies as imaging agents and drug conjugates have shown great potential for cancer diagnostics and therapeutics. However, site-specific modification of a nanobody with microbial transglutaminase (mTGase) encounters problems in protein separation and purification. Here, we describe a facile yet reliable strategy of immobilizing mTGase onto magnetic beads for site-specific nanobody modification. The mTGase immobilized on magnetic beads (MB-mTGase) exhibits catalytic activity nearly equivalent to that of the free mTGase, with good reusability and universality. Magnetic separation simplifies the protein purification step and reduces the loss of nanobody bioconjugates more effectively than size exclusion chromatography. Using MB-mTGase, we demonstrate site-specific conjugation of nanobodies with fluorescent dyes and polyethylene glycol molecules, enabling targeted immunofluorescence imaging and improved circulation dynamics and tumor accumulation in vivo. The combined advantages of MB-mTGase method, including high conjugation efficiency, quick purification, less protein loss, and recycling use, are promising for site-specific nanobody functionalization and biomedical applications.

Konjac glucomannan-based highly antibacterial active films loaded with thyme essential oil through bacterial cellulose nanofibers/Ag nanoparticles stabilized Pickering emulsions.

The aim of this study was to develop novel konjac glucomannan (KGM)-based highly antibacterial active films, where five types of films were prepared and compared. The microstructure results showed that KGM-based films loaded with thyme essential oil (TEO) through bacterial cellulose nanofibers/Ag nanoparticles (BCNs/Ag nanoparticles) stabilized Pickering emulsions (Type V films) displayed the smoothest surface and the most evenly dispersed TEO droplets as compared with the other four types of films. Moreover, Type V films showed the highest contact angle value (86.28°), the best thermal stability and mechanical properties. Furthermore, Type V films presented the highest total phenol content (13.23 mg gallic acid equivalent/g film) and the best antioxidant activity (33.96 %) as well as the best sustained-release property, thus showing the best antibacterial activity, which was probably due to that BCNs/Ag nanoparticles and TEO displayed a synergistic effect to some extent. Consequently, Type V film-forming solutions were used as coatings for tangerines. The results showed that the tangerines treated with Type V coatings displayed excellent fresh-keeping properties. Therefore, the coatings, KGM-based film-forming solutions loaded with TEO through BCNs/Ag nanoparticles stabilized Pickering emulsions, have great potential for the preservation of fruits and vegetables.

Synthesis of bacterial cellulose nanofibers/Ag nanoparticles: Structure, characterization and antibacterial activity.

The aim of this study was to compare the characterization of bacterial cellulose nanofibers/Ag nanoparticles (BCNs/Ag nanoparticles) obtained by three different pretreatment methods of BCNs (no pretreatment, sodium hydroxide activation pretreatment and TEMPO-mediated oxidation pretreatment), which were recoded as N-BCNs/Ag nanoparticles, A-BCNs/Ag nanoparticles and O-BCNs/Ag nanoparticles, respectively. The results of scanning electron microscopy and transmission electron microscopy showed the prepared Ag nanoparticles by three different pretreatment methods were spherical and dispersed on the surface of BCNs, while the Ag nanoparticles in O-BCNs/Ag nanoparticles displayed the smallest diameter with a value of 20.25 nm and showed the most uniform dispersion on the surface of BCNs. The ICP-MS result showed O-BCNs/Ag nanoparticles had the highest content of Ag nanoparticles with a value of 2.98 wt%, followed by A-BCNs/Ag nanoparticles (1.53 wt%) and N-BCNs/Ag nanoparticles (0.84 wt%). The cytotoxicity assessment showed that the prepared BCNs/Ag nanoparticles were relatively safe. Furthermore, the O-BCNs/Ag nanoparticles had the best antioxidant and antibacterial activities as compared with the other two types of BCNs/Ag nanoparticles, where O-BCNs/Ag nanoparticles destroyed the structure of bacterial cell membranes to lead the leakage of intracellular components. This study showed that O-BCNs/Ag nanoparticles as antibacterial agents have great potential in food packaging.