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| 001 | 614232 | ||
| 005 | 20250625124227.0 | ||
| 024 | 7 | _ | |a 10.3390/pharmaceutics15010225 |2 doi |
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| 100 | 1 | _ | |a Xu, Maokai |0 0000-0002-6267-3070 |b 0 |e Corresponding author |
| 245 | _ | _ | |a Dimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular Barriers |
| 260 | _ | _ | |a Basel |c 2023 |b MDPI |
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| 520 | _ | _ | |a Receptor-mediated transcytosis is an elegant and promising strategy for drug delivery across biological barriers. Here, we describe a novel ligand–receptor pair based on a dimeric, engineered derivative of the Pseudomonas aeruginosa lectin LecA, here termed Di-LecA, and the host cell glycosphingolipid Gb3. We characterized the trafficking kinetics and transcytosis efficiencies in polarized Gb3-positive and -negative MDCK cells using mainly immunofluorescence in combination with confocal microscopy. To evaluate the delivery capacity of dimeric LecA chimeras, EGFP was chosen as a fluorescent model protein representing macromolecules, such as antibody fragments, and fused to either the N- or C-terminus of monomeric LecA using recombinant DNA technology. Both LecA/EGFP fusion proteins crossed cellular monolayers in vitro. Of note, the conjugate with EGFP at the N-terminus of LecA (EGFP-LecA) showed a higher release rate than the conjugate with EGFP at the C-terminus (LecA-EGFP). Based on molecular dynamics simulations and cross-linking studies of giant unilamellar vesicles, we speculate that EGFP-LecA tends to be a dimer while LecA-EGFP forms a tetramer. Overall, we confidently propose the dimeric LecA chimeras as transcytotic drug delivery tools through Gb3-positive cellular barriers for future in vivo tests.Keywords:Gb3-binding lectin; glycosphingolipid; transcytosis; drug carrier; protein engineering; protein structure; valency; accelerated molecular dynamics |
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| 700 | 1 | _ | |a Antonova, Maria |b 1 |
| 700 | 1 | _ | |a Salavei, Pavel |b 2 |
| 700 | 1 | _ | |a Illek, Katharina |b 3 |
| 700 | 1 | _ | |a Meléndez, Ana Valeria |b 4 |
| 700 | 1 | _ | |a Omidvar, Ramin |0 0000-0003-1170-1382 |b 5 |
| 700 | 1 | _ | |a Thuenauer, Roland |0 P:(DE-H253)PIP1087856 |b 6 |
| 700 | 1 | _ | |a Makshakova, Olga |0 0000-0002-0615-3513 |b 7 |
| 700 | 1 | _ | |a Roemer, Winfried |0 P:(DE-H253)PIP1108890 |b 8 |e Corresponding author |
| 773 | _ | _ | |a 10.3390/pharmaceutics15010225 |g Vol. 15, no. 1, p. 225 - |0 PERI:(DE-600)2527217-2 |n 1 |p 225 |t Pharmaceutics |v 15 |y 2023 |x 1999-4923 |
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