title: Strategies and limitations in dendrimeric immunogen synthesis. The influenza virus M2e epitope as a case study.
abstract: Dendrimeric platforms such as multiple antigen peptides (MAPs) are regarded as one of the most efficacious approaches for antigenic presentation. Originally described as available by stepwise solid-phase peptide synthesis (SPPS), MAPs have also been prepared by chemical (thioether, oxime, hydrazone) ligation of appropriately functionalized tetra- or octavalent polylysine scaffolds with the peptide antigen to be multiply displayed. In this work, the advantages and limitations of two of the most frequent methods of MAP preparation, namely, chemoselective thioether ligation in solution, and all-solid-phase synthesis, have been tested in the case of a particularly troublesome epitope model, the ectodomain of protein M2 from influenza virus (M2e). The strong tendency of M2e to self-associate is a serious inconvenient for conjugation in solution, which as a result fails to produce the target MAPs with the specified number of M2e copies. In contrast, the fully stepwise SPPS approach is shown to be quite practical, especially when 6-aminohexanoic acid spacer units providing increased internal flexibility are inserted at each branching point.
authors.affiliation: Department of Experimental and Health Sciences, Pompeu Fabra University, Barcelona Biomedical Research Park, 08003 Barcelona, Spain.
authors.forename: Wioleta
authors.initials: W
authors.lastname: Kowalczyk
authors.forename: Beatriz G
authors.initials: BG
authors.lastname: de la Torre
authors.forename: David
authors.initials: D
authors.lastname: Andreu
mesh_terms: D000595:Amino Acid Sequence; D015119:Aminocaproic Acid / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000737:chemistry / Q000276:immunology; D050091:Dendrimers / Q000138:chemical synthesis* / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D008969:Molecular Sequence Data; D012694:Serine / Q000737:chemistry / Q000276:immunology; D012996:Solutions / Q000737:chemistry; D013440:Sulfides / Q000737:chemistry / Q000494:pharmacology; D014614:Vaccines, Synthetic / Q000737:chemistry* / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000145:classification / Q000276:immunology*; D000086082:Viroporin Proteins
chemical_list: D015119:Aminocaproic Acid; D000911:Antibodies, Monoclonal; D003545:Cysteine; D050091:Dendrimers; D000939:Epitopes; D012694:Serine; D012996:Solutions; D013440:Sulfides; D014614:Vaccines, Synthetic; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins