bc5b00547_si_001.pdf (1.41 MB)
Synthesis of Mannosylated Lipopeptides with Receptor Targeting Properties
journal contribution
posted on 2016-01-06, 00:00 authored by Bita Sedaghat, Rachel J. Stephenson, Ashwini
Kumar Giddam, Sharareh Eskandari, Simon H. Apte, David J. Pattinson, Denise L. Doolan, Istvan TothPresent on the surface of antigen
presenting cells (APCs), the
mannose receptor (MR) has long been recognized as a front-line receptor
in pathogen recognition. During the past decade many attempts have
been made to target this receptor for applications including vaccine
and drug development. In the present study, a library of vaccine constructs
comprising fluorescently labeled mannosylated lipid-dendrimers that
contained the ovalbumin CD4+ epitope, OVA323–339, as the model peptide antigen were synthesized using fluorenylmethyloxycarbonyl
(Fmoc) solid phase peptide synthesis (SPPS). The vaccine constructs
were designed with an alanine spacer between the O-linked mannose
moieties to investigate the impact of distance between the mannose
units on receptor-mediated uptake and/or binding in APCs. Uptake studies
performed on F4/80+ and CD11c+ cells showed
significant uptake and/or binding for lipopeptides containing mannose,
and also the lipopeptide without mannose when compared to the control
peptides (peptide with no lipid and peptide with no mannose and no
lipid). Furthermore, mannan inhibition assays demonstrated that uptake
of the mannosylated and lipidated peptides was receptor mediated.
To address the specificity of receptor uptake, surface plasmon resonance
studies were performed using biacore technology and confirmed high
affinity of the mannosylated and lipidated vaccine constructs toward
the MR. These studies confirm that both mannose and lipid moieties
play significant roles in receptor-mediated uptake on APCs, potentially
facilitating vaccine development.
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mannose unitsbiacore technologydrug developmentOVAAPCSPPSUptake studiesphase peptide synthesislipidated peptidescontrol peptidesmodel peptide antigenMRalanine spacerMannosylated Lipopeptidessurface plasmon resonance studiesmannosylatedlipid moietiesmannan inhibition assayspathogen recognitionreceptor uptakeReceptor Targeting Propertiesvaccine developmentCD 11c cellslipidated vaccinemannose receptor
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