Biology subjects

Walters, A.

Publications and source records attributed to Walters, A..

3 recordsLinked to original sources

bioRxiv2026

Extrahepatic, cell-specific delivery of LNPs through competitive inhibition of ApoE-mediated uptake

Targeted lipid nanoparticles (LNPs) for extrahepatic drug delivery are limited by apolipoprotein E (ApoE)-mediated hepatic accumulation. We developed NanoPilot, a modular fusion protein platform comprising antibodies and anchors blocking the low density lipoprotein receptor (LDLR) ApoE binding site, to block LNP liver uptake and redirect to target cells. NanoPilot can be applied to preformulated LNPs in 10 minutes with two pipetting steps. In vitro, an anti-CD3{varepsilon} NanoPilot increased T-cell transfection 40-fold and reduced monocyte transfection 10-fold in human peripheral blood mononucleocytes. In immunocompetent mouse models, NanoPilot-coated LNPs achieved 30-40% splenic and hepatic T-cell transfection whilst bulk liver accumulation was reduced 3-fold. An anti-c-Kit NanoPilot was further shown to enhance delivery to a haematopoietic stem cell-like cell line in an in-vitro co-culture assay. NanoPilot establishes a versatile framework for the systemic delivery of genetic therapies through concomitant cell-specific targeting and off-target blocking. Further research will assess potential clinical applications.

biochemistry↗

bioRxiv2026

Precise measurement of rodent drinking using CLiQR (Capacitive Lick Quantification in Rodents)

Accurate quantification of rodent licking behavior is essential for studies of fluid intake, including investigations of alcohol use disorder and obesity. Lickometry systems vary widely in sensing modality, cost, scalability, and data resolution. Also, many available systems require specialized housing or store only binary lick/no lick data based on thresholding. Here, we present CLiQR (Capacitive Lick Quantification in Rodents), an open-source capacitive lickometry system designed for high-throughput recording of licking behavior in home-cage environments while preserving the full capacitance time series. The system uses MPR121 capacitive sensors connected to custom metal-tipped serological pipette sippers and a centralized desktop computer to record data from up to 24 mice concurrently, with capacity for two-bottle choice experiments. Validation experiments demonstrated that the capacitive signals reliably distinguish licking from non-licking interactions. Total lick counts showed a positive correlation with measured fluid consumption (R2 = 0.849, p < 0.0001), confirming that detected events provide a meaningful proxy for intake. All information necessary to reproduce the system is shared openly in this manuscript and online. By combining scalability, full-trace data acquisition, and low cost, CLiQR provides a flexible and extensible platform for high-throughput behavioral neuroscience experiments and enables retrospective improvement of lick-detection algorithms.

animal behavior and cognition↗

bioRxiv2026

Bilayer acoustic force spectroscopy (BAFS) for quantifying receptor-antigen binding strength in immune synapses

Immune cell receptor - ligand interactions are key to cancer immunotherapy. However, receptor-ligand affinities often fail to predict T-cell mediated cancer killing, while immune-target cell binding strength measurements are limited by low precision and high non-specific binding. Here we present bilayer acoustic force spectroscopy (BAFS), a method to quantify the binding strength of receptors in immune synapses that virtually eliminates non-specific binding and increases the resolving power by up to 50-fold. By replacing target cells with a supported lipid bilayer functionalized with antigens, BAFS avoids antigen-independent interactions and target cell heterogeneity, while maintaining the spatial self-organization of receptors that typifies active immune synapses. We demonstrate the high sensitivity and control by showing how CAR T-cell synapse strength depends on CD19 antigen density, and by revealing that CD8 synergistically strengthens {beta}TCR-pMHC synapses independently of Lck recruitment to CD8. BAFS is a general method that can be used broadly in immunotherapy screening and to dissect the complex molecular interactions that underpin immune synapse activation.

biophysics↗