Similar Datasets
Project description:To understand the molecular determinants of B12(VHH)-CAR-T polyfunctionality, we simultaneously measured the protein and RNA expression levels of 95 single CD4+ B12(VHH)-CAR-T cells isolated from IMR5 tumor-bearing mouse and re-stimulated with IMR5 tumor cells in vitro. We identified two CAR-T clusters, low polyfunctionality and high polyfunctionality subsets, in a 2D t-SNE plot. Moreover, our single-cell mRNA expression profiling revealed 32 genes that displayed statistically significant, concordant differences between the two cell subsets.
2023-08-17 | GSE221411 | GEO
Project description:Additive effects of TNF and IL-6 were evaluated in cell-based model for rheumatoid arthritis (RA) and then compared to treatment with anti-TNF/IL-6 NANOBODY® VHH, Humira and Sylvant by RNA-seq
2022-12-31 | GSE200291 | GEO
Project description:SUMOylation is a post-translational modification regulating protein localization, stability, and activity, with effects varying depending on the identity of the conjugated SUMO protein (SUMO1 or SUMO2/3) and type of SUMOylation (mono-, multi-, or poly-). We previously developed a small 32 amino acids SUMOylation tag derived from the ZNF451 E3 ligase (hereafter “ZNF”) that biases substrates toward SUMO2/3 conjugation and modulates the transcriptional activity of p53. To determine how enhanced SUMOylation affects protein localization, we fused ZNF to the GFP-binding nanobody vhhGFP4 (VHH), creating VHH-ZNF to drive SUMOylation of GFP-tagged substrates in trans. In vitro, VHH-ZNF increased SUMO2/3 modification of p53-GFP, preferentially generating polySUMO2 chains at the canonical K386 site; using a chain-deficient SUMO2(K0) shifted the pattern toward multi-SUMOylation. VHH-ZNF did not modify HA-p53 alone but SUMOylated it in the presence of p53-GFP, consistent with proximity-based modification within p53 tetramers. In HEK293 cells co-expressing p53-GFP and VHH-ZNF, immunoblotting and proteomics confirmed increased SUMO2/3 conjugation at K386 that was abrogated by the E1 inhibitor ML792. Fluorescence microscopy analyses revealed that SUMOylated p53 transitions from a diffuse nuclear distribution to SUMO-positive nuclear foci that partially overlap with promyelocytic leukemia (PML) nuclear bodies. Overall, we developed a new method to increase the SUMOylation of GFP-tagged proteins and to visualize SUMOylation-dependent relocalization in cells.
2026-03-24 | PXD069854 | Pride
Project description:Proteomics profiles of extracellular vesicles isolated using a new cryogel-based immunoaffinity chromatography system functionalized with single-domain VHH antibodies for selective isolation of EVs from human plasma.
2025-12-08 | PXD070239 | Pride
Project description:Camelids are capable of producing both conventional tetrameric antibodies (Abs) and dimeric heavy-chain antibodies (HCAbs). While B cells generating these two types of Abs exhibit distinct B-cell receptors (BCRs), it remains unclear whether these two B cell populations differ in their phenotypes and developmental processes. Here, we collected eight PBMC samples before and after immunization from four Bactrian camels and conducted single-cell 5’ RNA sequencing. We characterized the functional subtypes and differentiation trajectories of circulating B cells in camels, including native B cells, memory B cells, intermediate B cells, atypical B cells, and plasma cells. Additionally, we reconstructed single-cell BCR sequences and revealed the IGHV and IGHC gene types. We found that B cells with variable genes of HACbs (VHH) were widely present in various functional subtypes and showed highly overlapping differentiation trajectories to B cells with variable genes of conventional Abs (VH). After immunization, the transcriptional changes in VHH+ and VH+ B cells were also largely consistent. Our study not only elucidates the cellular context of HCAb production in camels, but also lays the foundation for the development of single B cell-based nanobody screening.
2024-04-15 | GSE238082 | GEO