Journal: bioRxiv
Article Title: CELLISA – a cell-cell binding assay for evaluation of nanovesicle targeting proteins
doi: 10.64898/2026.04.09.717595
Figure Lengend Snippet: (A) Cartoon describing the targets, binder types, and linkers used in this investigation. (B) sCELLISA data identifying candidate binders to Jurkat T cells. Each symbol represents the average of two fields of view from a biological replicate (n = 3), the bar represents the mean, and the error bars represent standard error of the mean. (C) Cartoon depicting the goal of evaluating sCELLISA hits via EV targeting. (D) Quantification of 15k (filled bars) and 120k (outlined bars) EVs targeted to Jurkat T cells with binders identified from the sCELLISA assay. The cellular fluorescence of each condition was background fluorescence subtracted (fluorescence attributable to cells only) and normalized by EV fluorescence determined by NanoFCM. Each bar graph represents an independent experiment, and each bar represents the mean of three biological replicates (n = 3). The error bars represent the standard error of the mean. To evaluate whether binders resulted in increased EV binding compared to the scaffold negative control, a two-way ANOVA was performed, and the Dunnett’s multiple comparison test is shown (****, p < 0.0001; ***, p < 0.001; **, p < 0.01; *, p < 0.05; ns, p > 0.05). (E) Cartoon depicting evaluation of delivery improvement by candidate binders via lentivirus. (F) Quantification of functional titers of unconcentrated lentiviruses expressing mutant VSV-G and the binder indicated on the x-axis. Filled and outlined bars each represent an independent experiment. Each bar represents the mean functional titer as determined via linear regression by calculating the multiplicity of infection using the percent of Jurkat cells expressing dsRedExpress2 (also see Methods: Lentiviral Delivery Studies). Error bars represent the standard error of the slope of the linear regression. To identify if a binder improved delivery compared to the scaffold negative control, a pairwise comparison of each slope from each linear regression to the slope of the scaffold using a one-sample t-test was performed (***, p < 0.001; **, p < 0.01; *, p < 0.05; ns, p > 0.05).
Article Snippet: The lentiviral transfer plasmid was grown in NEB Stable competent E. coli (New England Biolabs C3040) and grown at 30 °C.
Techniques: Fluorescence, Binding Assay, Negative Control, Comparison, Functional Assay, Expressing, Mutagenesis, Infection
