TY - JOUR
AU - Xu, Maokai
AU - Antonova, Maria
AU - Salavei, Pavel
AU - Illek, Katharina
AU - Meléndez, Ana Valeria
AU - Omidvar, Ramin
AU - Thuenauer, Roland
AU - Makshakova, Olga
AU - Roemer, Winfried
TI - Dimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular Barriers
JO - Pharmaceutics
VL - 15
IS - 1
SN - 1999-4923
CY - Basel
PB - MDPI
M1 - PUBDB-2024-05772
SP - 225
PY - 2023
AB - 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
LB - PUB:(DE-HGF)16
C6 - pmid:36678854
UR - <Go to ISI:>//WOS:000918781700001
DO - DOI:10.3390/pharmaceutics15010225
UR - https://bib-pubdb1.desy.de/record/614232
ER -