1. Sortase-A mediated chemoenzymatic lipidation of single-domain antibodies for cell membrane engineering
- Author
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Steffen Wöll, Lee Kim Swee, Regina Scherließ, Christopher Bachran, and Stefan Schiller
- Subjects
T-Lymphocytes ,Cell ,Pharmaceutical Science ,Lipid-anchored protein ,02 engineering and technology ,030226 pharmacology & pharmacy ,Polyethylene Glycols ,Cell membrane ,Cell therapy ,Mice ,03 medical and health sciences ,Drug Delivery Systems ,0302 clinical medicine ,Bacterial Proteins ,medicine ,Animals ,Cells, Cultured ,Liposome ,Chemistry ,Cell Membrane ,General Medicine ,Single-Domain Antibodies ,Aminoacyltransferases ,021001 nanoscience & nanotechnology ,Lipids ,Cell biology ,Mice, Inbred C57BL ,Cysteine Endopeptidases ,medicine.anatomical_structure ,Targeted drug delivery ,Sortase A ,Liposomes ,Drug delivery ,0210 nano-technology ,Biotechnology - Abstract
Purpose Membrane engineering has versatile applications in adoptive cell therapies, immune therapy or drug delivery. Incorporation of lipidated antibody-derived ligands into cells may enforce supraphysiological cell interactions that offer new therapeutic approaches. A challenge is the defined synthesis of lipidated ligands that effectively interact with such membranes. Methods Sortase-A was used to attach a PEGylated, dimyristyl lipid-anchor on single-domain antibodies (VHH). The membrane insertion was investigated on liposomal bilayers, myeloid-derived suppressor cells (MDSC) and T cells. Results The lipidated VHHs remodeled liposomal as well as cellular membranes. The VHH carrying liposomes were successfully targeted towards antigen-positive cells. MDSC and T cells were both modified with lipidated VHHs as detected with an FITC-anti-llama antibody. T cells that carried an anti-CD11b VHH showed cellular association in vitro with CD11b+Gr-1+ MDSC in a two-dimensional magnetic activated cell sorting / flow-cytometry assay. Conclusion The applied combination of chemoenzymatic ligation, PEGylated lipid anchors and single-domain antibodies delivers water-soluble and chemically defined lipidated ligands, which readily associate with liposomal and cellular membranes. This enables liposomal drug targeting and artificial cell-cell interactions. Hence, the presented concept for lipidation of single-domain antibodies is promising for further application in the field of drug delivery or cell-based therapies.
- Published
- 2020