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000614232 1001_ $$00000-0002-6267-3070$$aXu, Maokai$$b0$$eCorresponding author
000614232 245__ $$aDimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular Barriers
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000614232 520__ $$aReceptor-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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000614232 7001_ $$aAntonova, Maria$$b1
000614232 7001_ $$aSalavei, Pavel$$b2
000614232 7001_ $$aIllek, Katharina$$b3
000614232 7001_ $$aMeléndez, Ana Valeria$$b4
000614232 7001_ $$00000-0003-1170-1382$$aOmidvar, Ramin$$b5
000614232 7001_ $$0P:(DE-H253)PIP1087856$$aThuenauer, Roland$$b6
000614232 7001_ $$00000-0002-0615-3513$$aMakshakova, Olga$$b7
000614232 7001_ $$0P:(DE-H253)PIP1108890$$aRoemer, Winfried$$b8$$eCorresponding author
000614232 773__ $$0PERI:(DE-600)2527217-2$$a10.3390/pharmaceutics15010225$$gVol. 15, no. 1, p. 225 -$$n1$$p225 $$tPharmaceutics$$v15$$x1999-4923$$y2023
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