Evaluation of asymmetric immunoliposomal nanoparticles for cellular uptake

Abstract

Effective and targeted in vivo delivery of polynucleotide therapeutics is the key for the treatment of many diseases. Asymmetric immunoliposomes can be used as vehicles to deliver polynucleotides effectively because the two leaflets of the bilayer can have different compositions, which enhance the delivery capacity. The formation and in vitro cellular uptake of asymmetric immunoliposomes containing polynucleotide cargoes were studied here. Maleimide functionalized DSPE-PEG (2000) were incorporated into the outer leaflet to produce asymmetric liposomes capable of covalently attaching antibodies. Thiolated antibodies from both human and rabbit origin were conjugated to produce asymmetric pendant-type immunoliposomes that retain their specificity towards detection antibodies through the formation process. Human IgG conjugated asymmetric immunoliposomes were readily internalized (> 20 per cell) by macrophage, HEPG2, and CV-1 monkey kidney cells. The cells internalized the liposomal nanoparticles by the endocytic pathway. The immunoliposome-encapsulated endosomes were intact for at least 5 days and sequestered the plasmid from expression by the cell.

Endothelial-Specific Targeting of RhoA Signaling via CD31 Antibody-Conjugated Nanoparticles

“…4A-B), and selected Lipofectamine 2000 for further studies. To evaluate the binding of the CD31FL particles on the murine CD31 expressing or control HEK293T cells, we detected the CD31 antibody presence by the specific binding of the specific FITC-conjugated secondary antibody (Whittenton, et al, 2013). The selective binding of the CD31FL liposomes upon CD31 expression was verified at all evaluated time points (0.5 h, 1 h, 4 h; Fig.…”

“…Generally, 10,000 events were acquired per sample (Gholizadeh, Dolman, et al, 2018). The data were further analyzed by FlowJo and expressed as mean fluorescence intensity (Gholizadeh, Visweswaran, et al, 2018;Whittenton, Pitchumani, Thevananther & Mohanty, 2013). The pcDNAI Neo mouse PECAM FL plasmid was a gift from Steven Albelda (Addgene plasmid # 42901 ; http://n2t.net/addgene:42901 ; RRID:Addgene_42901).…”

Asymmetric Lipid Vesicles: Techniques, Applications, and Future Perspectives as an Innovative Drug Delivery System

“…This methodology has proven useful; for example, Whittenton et al used this technique to generate immunoliposomes with DOTAP and DMPC in the inner leaflet and DMPC, POPC, NBD(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl)-PC, and NBD-PS in the outer leaflet, encapsulating polynucleotides to evaluate the cellular uptake [ 27 ]. Additionally, de Matos et al synthesized stable asymmetric liposomes of around 200 nm made from DOTAP and DSPC with a simple centrifugation-based w / o emulsion capable of entrapping pDNA [ 25 ].…”

Leveraging immunoliposomes as nanocarriers against SARS-CoV-2 and its emerging variants

“…Maleimide thiol chemistry has also been used for normal cellular uptake studies [81] . In creation of asymmetric immunoliposomes that maintain their affinity and avidity during the development process, antibodies from both human and rabbit were thiolated and attached to maleimide-functionalized liposomes.…”