Dataset Information

MudPIT analyses of the human proteins biotinylated by proximity to exogenously expressed biotin ligase (BioID)

ABSTRACT: To identify non-specific biotinylation events, HEK293 cells transfected with a plasmid carrying only the biotin ligase moiety were used as negative controls to BioID experiments. Cells were lysed in the presence of detergents. After purification of biotinylated proteins on streptavidin-conjugated beads, the eluates were precipitated with TCA. After washing with acetone, the protein mixtures were digested with endoproteinase Lys-C and trypsin (Promega) and analyzed by MudPIT.

INSTRUMENT(S): LTQ

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Laurence Florens

PROVIDER: MSV000080809 | MassIVE | Thu Mar 30 11:44:00 BST 2017

SECONDARY ACCESSION(S): PXD006222

REPOSITORIES: MassIVE

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Publications

MVA E2 therapeutic vaccine for marked reduction in likelihood of recurrence of respiratory papillomatosis.

Cabo Beltran Olga Rodriguez OR Rosales Ledezma Ricardo R

Head & neck 20190103 3

<h4>Background</h4>Recurrent respiratory papillomatosis (RRP) or laryngeal papillomatosis is a disease caused by papillomavirus infection.<h4>Methods</h4>In this phase I/II clinical trial, we evaluated the efficacy of the modified vaccinia Ankara (MVA) E2 virus in the treatment of RRP. Twenty-nine patients (18 female and 11 male) underwent injection of MVA E2 directly into the borders of the vocal cords where lesions were seen and were monitored by direct laryngoscopy. The immune response was as ...[more]

PMID: 30605254

Similar Datasets

Project description:Abstract: Controlled release of immunomodulatory cytokines like interleukin-4 (IL-4) from biomaterials can modulate host immune cell behavior to improve tissue integration, however options for modifying complex biomaterials remain limited. Bioconjugation strategies using biotin-avidin interactions offer a promising approach since biotin can be conjugated to proteins, biomaterials, and even cells, without compromising their function. Although it is known that conjugation of biotin to large biomolecules reduces its binding affinity for avidin and avidin variants, the potential to control release of biotinylated molecules via leveraging affinity interactions has not been thoroughly investigated. Moreover, the effects of biotin and avidin variants on innate immunity are poorly understood. Therefore, the goal of this study was to determine if biotin-avidin interactions could be manipulated to control the release of IL-4 from biomaterials and to investigate the subsequent effects on primary human macrophage phenotype. First, we characterized the effects of soluble biotin, avidin, and avidin variants, streptavidin and CaptAvidin, on the phenotype of primary human macrophages using RNA sequencing and found that CaptAvidin significantly influenced macrophage gene expression while other variants did not. Then, after evaluating how bioconjugation parameters influenced biotin density and avidin variant binding to porous gelatin scaffolds, we found that biotin-avidin affinity interactions influenced the spontaneous release of biotinylated IL-4 (bIL-4) from biotinylated and desthiobiotinylated scaffolds bound with either avidin or streptavidin for up to ten days in vitro. Finally, we measured the response of primary human macrophages (8 donors) cultured on modified scaffolds, finding an increase in IL-4 driven genes when bIL-4 was released via biotin-streptavidin affinity interactions compared to desorption-based release. These results highlight how biotin-avidin interactions can be leveraged for spontaneous release to achieve an immunomodulatory drug delivery system.