We utilize a plane-wave density-functional theory approach to investigate the vibrational properties of the all-trans ferroelectric phase of poly͑vinylidene fluoride͒ ͑␤-PVDF͒ showing that its stable state corresponds to the Ama2 structure with ordered dihedral tilting of the VDF monomers along the polymer chains. We then combine our theoretical analysis with IR spectroscopy to examine vibrations in oligomer crystals that are structurally related to the ␤-PVDF phase. We demonstrate that these materials-which can be grown in a highly crystalline form-exhibit IR activity similar to that of ␤-PVDF, making them an attractive choice for the studies of electroactive phenomena and phase transitions in polymer ferroelectrics.

Broadening gene therapy applications requires manufacturable vectors that efficiently transduce target cells in humans and preclinical models. Conventional selections of adeno-associated virus (AAV) capsid libraries are inefficient at searching the vast sequence space for the small fraction of vectors possessing multiple traits essential for clinical translation. Here, we present Fit4Function, a generalizable machine learning (ML) approach for systematically engineering multi-trait AAV capsids. By leveraging a capsid library that evenly samples the manufacturable sequence space, reproducible screening data are generated to train accurate sequence-to-function models. Combining six models, we designed a multi-trait (liver-targeted, manufacturable) capsid library and validated 89% of library variants on all six predetermined criteria. Furthermore, the models, trained only on mouse in vivo and human in vitro Fit4Function data, accurately predicted AAV capsid variant biodistribution in macaque. Top candidates exhibited high production yields, efficient murine liver transduction, up to 1000-fold greater human hepatocyte transduction, and increased enrichment, relative to AAV9, in a screen for liver transduction in macaques. The Fit4Function strategy ultimately makes it possible to predict cross-species traits of peptide-modified AAV capsids and is a critical step toward assembling an ML atlas that predicts AAV capsid performance across dozens of traits.

Vibrational spectroscopy is a powerful analytical tool that assesses molecular properties based on spectroscopic signatures. In this study, the effect of gold nanoparticle morphology (spherical vs multi-branched) was assessed for the characterization of a Raman signal (ie, molecular fingerprint) that may be helpful for numerous medical applications. Multi-branched gold nanoparticles (MBAuNPs) were fabricated using a green chemistry method which employed the reduction of gold ion solute by 2-[4-(2-hydroxyethyl)-1-piperazyl] ethane sulfonic acid. Two types of reporter dyes, indocyanine (IR820 and IR792) and carbocyanine (DTTC [3,3′-diethylthiatricarbocyanine iodide] and DTDC [3,3′-diethylthiadicarbocyanine iodide]), were functionalized to the surface of the MBAuNPs and stabilized with denatured bovine serum albumin, thus forming the surface-enhanced Raman spectroscopy tag. Fluorescein isothiocyanate-conjugated anti-epidermal growth factor receptor to the surface-enhanced Raman spectroscopy tags and the properties of the resulting conjugates were assessed through determination of the Raman signal. Using the MBAuNP Raman probes synthesized in this manner, we demonstrated that MBAuNP provided significantly more surface-enhanced Raman scattering signal when compared with the associated spherical gold nanoparticle of similar size and concentration. MBAuNP enhancements were retained in the surface-enhanced Raman spectroscopy tags complexed to anti-epidermal growth factor receptor, providing evidence that this could be a useful biological probe for enhanced Raman molecular fingerprinting. Furthermore, while utilizing IR820 as a novel reporter dye linked with MBAuNP, superior Raman signal fingerprint results were obtained. Such results provide significant promise for the use of MBAuNP in the detection of numerous diseases for which biologically specific surface markers exist.

No abstract

Surface enhanced Raman scattering (SERS) is a sensitive and reproducible vibrational spectroscopic technique used to detect and characterize molecules near the surface of noble metals like Au, Ag, Pt, Cu, etc. SERS enhances Raman signals through light-induced plasmonic vibrations occurring on irregular metal surfaces and localized electromagnetic augmentation. To better define nano-scale regions of the Raman signal enhancement, we generated gold nanoparticles with a unique multi-branched configuration along with surface-adsorbed fluorescent reporter molecules. The reporter molecules included a set of near-infra red active fluorescent dyes IR820 (green cyanine, photo electronic dye), DTTC (3, 3'-diethylthiatricarbocyanine iodide) and DTDC (3, 3'-diethylthiadicarbocyanine iodide). We employed a one-pot synthesis method in order to generate a stellate configuration in gold nanoparticles through the reduction of HAuCl4 with Good's buffer, HEPES, at pH 7.4 and room temperature. A cell viability assay was performed with normal esophageal cells exposed to the multi-branched gold nanoparticles and SERS molecules to assess their toxicity. Our results demonstrate the capacity of multibranched gold nanoparticles linked to Raman reporter molecules to generate distinct signature spectra and, with the exception of the gold nanoparticles functionalized with DTTC, remain non-toxic to normal esophageal cells.