Construction and validation of a synthetic phage-displayed nanobody library.

Nanobodies derived from camelids and sharks offer unique advantages in therapeutic applications due to their ability to bind to epitopes that were previously inaccessible. Traditional methods of nanobody development face challenges such as ethical concerns and antigen toxicity. Our study presents a synthetic, phagedisplayed nanobody library using trinucleotide-directed mutagenesis technology, which allows precise amino acid composition in complementarity-determining regions (CDRs), with a focus on CDR3 diversity. This approach avoids common problems such as frameshift mutations and stop codon insertions associated with other synthetic antibody library construction methods. By analyzing FDA-approved nanobodies and Protein Data Bank sequences, we designed sub-libraries with different CDR3 lengths and introduced amino acid substitutions to improve solubility. The validation of our library through the successful isolation of nanobodies against targets such as PD-1, ATXN1 and STAT3 demonstrates a versatile and ethical platform for the development of high specificity and affinity nanobodies and represents a significant advance in biotechnology.

Complete mitochondrial genome of the crinoid Poliometra prolixa (Crinoidea: Comatulida: Antedonidae).

Poliometra prolixa Sladen, 1881, is a comatulid crinoid found in the Arctic deep sea. In this study, we report the complete mitochondrial genome sequence of P. prolixa (Comatulida: Antedonidae). The complete mitogenome of P. prolixa was 15,916 bp long and comprised 13 protein-coding genes (PCGs), 22 transfer RNA genes, and 2 ribosomal RNA genes. The base composition of the P. prolixa mitogenome was 24.0% A, 44.9% T, 19.0% G, and 12.1% C. Phylogenetic analysis using all PCGs of the complete mitogenome confirmed the inclusion of P. prolixa within the Comatulida.

Validation of Direct Boiling Method for Simple and Efficient Genomic DNA Extraction and PCR-based Macroalgal Species Determination.

A rapid, simple, and cost-effective total DNA extraction method referred to as a direct boiling method for PCR-based algal species determination was validated using representative species of green, brown, and red algae. Each sample was briefly minced in two buffers, Tris-EDTA (TE) or PCR buffer, and transferred to a heat block for boiling at 98°C for 5 min. No detergent was used in this experiment. The entire DNA isolation procedure was completed within 10 min. After brief centrifugation, the supernatant was directly used as a template for PCR. As a result, all genomic DNA markers were successfully amplified and sequenced from each algal taxon. Regardless of DNA quality, the direct boiling method is very suitable to identify unknown species of algae from a large amount of samples in a limited time and can be applied broadly to routine seasonal or annual monitoring of algae. DNA from an Undaria pinnatifida sporophyte, however, was not successfully extracted by this direct boiling method, probably due to a high concentration of polysaccharides.

Synthesis and characterization of poly(vinylpyrrolidone)-capped silicon-nanoparticles.

Polymer-capped silicon nanoparticles (Si NPs) were prepared using poly(vinylpyrrolidone) (PVP) as a protection layer to minimize the surface oxidation of the surface. The wet synthetic procedure were performed at room temperature via a one-pot synthesis. The PVP capped Si-NPs were characterized by thermo gravimetric analysis (TGA), Fourier transform infrared spectroscopy (FT-IR), high resolution transmission electron microscopy (HR-TEM) and field emission scanning electron microscopy (FE-SEM). HR-TEM micrograms confirm the presence of crystalline Si-NPs. The pattern by Selected Area Electron Diffraction (SAED) reveals (111), (220) lattice planes which are consistent with the cubic-structured crystalline silicon. The mean size of Si-NPs is estimated to be ca. 10 nm, which is larger than those from conventional wet synthetic methods for silicon quantum dots.