Biology subjects

Zohar, O.

Publications and source records attributed to Zohar, O..

2 recordsLinked to original sources

bioRxiv2026

cloneXplorer: A high-throughput clone discovery platform based on conical microwell arrays

Antigen-specific T cell populations are of great value for studying immune recognition but tedious to generate by limiting dilution or cloning. Here, we develop a streamlined approach to generate antigen-specific T cell clones directly from peripheral blood using the cloneXplorer, a live-cell analysis and clone isolation platform based on conical microwell arrays. This platform continuously monitors cell proliferation, cytokine secretion, and surface markers in up to 100,000 single cell co-cultures, enabling the identification of rare, functionally defined T cells, which can be recovered for clonal expansion or sequence analysis. We benchmark the platform by performing several key demonstrations. First, we show that this platform can efficiently generate monoclonal cell populations from cell lines and human T cells. Next, we demonstrate that antigen-specificity can be identified at single cell resolution using a co-culture of Jurkat cells expressing NFAT-GFP, CD8, and a T cell receptor and K562 antigen presenting cells (APC) expressing a peptide library. Thereafter, we show that immune activation in mouse and human primary samples can be monitored by time lapse analysis of Interferon gamma (IFN-{gamma}) secretion in individual microwell co-cultures using a fluorescent sandwich assay. Finally, we combine these capabilities in a proof-of-concept demonstration, which uses IFN-{gamma} secretion and the presence of CD8 surface markers as hierarchical gates to isolate and expand antigen-specific T cells from human peripheral blood, and we verify their specificity by tetramer staining. Together, these results showcase potential applications of the cloneXplorer platform in cell line development, and in screening and validating immune receptor interactions with specific antigens. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=156 SRC="FIGDIR/small/699323v1_ufig1.gif" ALT="Figure 1"> View larger version (48K): org.highwire.dtl.DTLVardef@1aafc29org.highwire.dtl.DTLVardef@91272dorg.highwire.dtl.DTLVardef@1a306eeorg.highwire.dtl.DTLVardef@1bfd54_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

bioRxiv2025

Nanobubble based sonobiopsy reveals circulating protein signatures of BBB opening and glioblastoma

Focused ultrasound (FUS) and microbubbles can transiently increase blood brain barrier (BBB) permeability, yet verification of BBB opening (BBBO) relies mainly on contrast-enhanced MRI, offering limited insight into the molecular consequences of barrier modulation. Sonobiopsy, which uses FUS induced BBBO to release brain derived molecules into the bloodstream, provides a molecular readout from blood samples. Nanobubbles (NBs) are smaller agents that circulate more effectively in the brain microvasculature and have shown enhanced BBBO in capillaries. Here, NB-mediated proteomic sonobiopsy is used to improve biomarker efflux and increase molecular sensitivity, with the goal of defining the molecular signature produced by BBBO in healthy and glioblastoma (GBM) models, and distinguishing biomarkers associated with tumor pathology from BBBO biomarkers. Plasma collected before and after NB-mediated FUS underwent data-independent acquisition-based mass spectrometry, revealing post-BBBO changes in healthy and tumor bearing mice. In healthy cohorts, 77 proteins were reproducibly altered after BBBO. Six proteins (Dpysl3, Myl1, Mybpc1, Vsig4, Krt33a, Krtap6-5) were detectable only after BBBO in healthy mice. In 005 glioma bearing mice, the BBBO signature identified in healthy animals was preserved, and comparison with matched shams isolated tumor specific effects. Three proteins, Itih4, Lrg1, and Hp, rose significantly after BBBO only in GBM and reached higher post treatment levels than in shams, nominating candidate GBM-associated markers accessible via blood. These findings establish NB-mediated proteomic sonobiopsy as a promising method for BBBO verification and for detecting GBM associated protein signals, supporting the development of scalable BBBO confirmation and protein-based diagnostics in neuro oncology. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=92 SRC="FIGDIR/small/692317v1_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@d0186aorg.highwire.dtl.DTLVardef@1f2fe8borg.highwire.dtl.DTLVardef@25921forg.highwire.dtl.DTLVardef@9c3566_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗