Synthesis,Solution Conformation,and Antibody Recognition of Oligotuftsin-Based Conjugates Containing a beta-Amyloid(4-10)Plaque-Specific Epitope

摘要

One possible therapeutic approach to treat or prevent Alzheimer's disease(AD)is immunotherapy.On the basis of the identification of Abeta(4-10)(FRHDSGY)as the predominant B-cell epitope recognized by therapeutically active antisera from transgenic AD mice,conjugates with defined structures containing the epitope peptide attached to a tetratuftsin derivative as an oligopeptide carrier were synthesized and their structure characterized.To produce immunogenic constructs,the Abeta(4-10)epitope alone or flanked by a-or beta-alanine residues was attached through an amide bond to the tetratuftsin derivative(Ac-[TKPKG]_4-NH_2)or to a carrier peptide elongated by a promiscuous T-helper cell epitope(Ac-FFLLTRILTIPQSLD-[TKPKG]_4-NH_2).The conformational preferences of the carrier and conjugates were examined by CD spectroscopy in water and in 1:1 and 9:1 TFE:water mixtures(v/v).We found that the presence of flanking dimers in the conjugates had no effects on the generally unordered solution conformation of the conjugates.However,conjugates with an elongated peptide backbone exhibited CD spectra indicative for a partially ordered secondary structure in the presence of TFE.Comparative ELISA binding studies,using monoclonal antibody raised against the beta-amyloid(1-17)peptide,showed that conjugates with T-helper cell epitope in the carrier backbone exhibited decreased monoclonal antibody recognition.However,we found that this effect was compensated in conjugates comprising the Abeta(4-10)B-cell epitope with the beta-alanine dimer flanking regions at both N-and C-termini.Results suggest that modification of the B-cell epitope peptide from Abeta with rational combination of structural elements(e.g.conjugation to carrier,introduction of flanking dimers)can result in synthetic antigen with preserved antibody recognition.