Synthesis, Solution Conformation, and Antibody Recognition of Oligotuftsin-Based Conjugates Containing a -Amyloid(4-1
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- 作者：Marilena Manea ; Ferenc Hudecz ; Michael Przybylski ; and Gá ; bor Mezõ
- 刊名：Bioconjugate Chemistry
- 出版年：2005
- 出版时间：July 2005
- 年：2005
- 卷：16
- 期：4
- 页码：921 - 928
- 全文大小：162K
- 年卷期：v.16,no.4(July 2005)
- ISSN：1520-4812
文摘
One possible therapeutic approach to treat or prevent Alzheimer's disease (AD) is immunotherapy.On the basis of the identification of A
(4-10) (FRHDSGY) as the predominant B-cell epitope recognizedby therapeutically active antisera from transgenic AD mice, conjugates with defined structurescontaining the epitope peptide attached to a tetratuftsin derivative as an oligopeptide carrier weresynthesized and their structure characterized. To produce immunogenic constructs, the A(4-10)epitope alone or flanked by 
- or -alanine residues was attached through an amide bond to thetetratuftsin derivative (Ac-[TKPKG]4-NH2) or to a carrier peptide elongated by a promiscuous T-helpercell epitope (Ac-FFLLTRILTIPQSLD-[TKPKG]4-NH2). The conformational preferences of the carrierand 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 generallyunordered solution conformation of the conjugates. However, conjugates with an elongated peptidebackbone exhibited CD spectra indicative for a partially ordered secondary structure in the presenceof TFE. Comparative ELISA binding studies, using monoclonal antibody raised against the -amyloid(1-17) peptide, showed that conjugates with T-helper cell epitope in the carrier backbone exhibiteddecreased monoclonal antibody recognition. However, we found that this effect was compensated inconjugates comprising the A(4-10) B-cell epitope with the -alanine dimer flanking regions at bothN- and C-termini. Results suggest that modification of the B-cell epitope peptide from A with rationalcombination of structural elements (e.g. conjugation to carrier, introduction of flanking dimers) canresult in synthetic antigen with preserved antibody recognition.
(4-10) (FRHDSGY) as the predominant B-cell epitope recognizedby therapeutically active antisera from transgenic AD mice, conjugates with defined structurescontaining the epitope peptide attached to a tetratuftsin derivative as an oligopeptide carrier weresynthesized and their structure characterized. To produce immunogenic constructs, the A(4-10)epitope alone or flanked by - or -alanine residues was attached through an amide bond to thetetratuftsin derivative (Ac-[TKPKG]4-NH2) or to a carrier peptide elongated by a promiscuous T-helpercell epitope (Ac-FFLLTRILTIPQSLD-[TKPKG]4-NH2). The conformational preferences of the carrierand 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 generallyunordered solution conformation of the conjugates. However, conjugates with an elongated peptidebackbone exhibited CD spectra indicative for a partially ordered secondary structure in the presenceof TFE. Comparative ELISA binding studies, using monoclonal antibody raised against the -amyloid(1-17) peptide, showed that conjugates with T-helper cell epitope in the carrier backbone exhibiteddecreased monoclonal antibody recognition. However, we found that this effect was compensated inconjugates comprising the A(4-10) B-cell epitope with the -alanine dimer flanking regions at bothN- and C-termini. Results suggest that modification of the B-cell epitope peptide from A with rationalcombination of structural elements (e.g. conjugation to carrier, introduction of flanking dimers) canresult in synthetic antigen with preserved antibody recognition.