期刊
BIOPOLYMERS
卷 104, 期 5, 页码 560-576出版社
WILEY
DOI: 10.1002/bip.22677
关键词
antibody recognition; synthetic antigenic probes; structural-activity relationship; surface plasmon resonance; antibody affinity
资金
- Laboratory of Peptide and Protein Chemistry and Biology of the University of Florence
- University P. et M. Curie. Fondazione Ente Cassa Risparmio di Firenze (Italy)
- Agence Nationale de la Recherche (France) [ANR-09-CEXC-013-01]
- Institute for Advanced Studies of University of Cergy-Pontoise
Antibody detection in autoimmune disorders, such as multiple sclerosis (MS) and Rett syndrome (RTT) can be achieved more efficiently using synthetic peptides. The previously developed synthetic antigenic probe CSF114(Glc), a type I' beta-turn N-glucosylated peptide structure, is able to recognize antibodies in MS and RTT patients' sera as a sign of immune system derangement. We report herein the design, synthesis, conformational analysis, and immunological evaluation of a collection of glycopeptide analogs of CSF114(Glc) to characterize the specific role of secondary structures in MS and RTT anti-body recognition. Therefore, we synthesized a series of linear and cyclic short glucosylated sequences, mimicking different b-turn conformations, which were evaluated in inhibition enzyme-linked immunosorbent assays (ELISA). Calculated IC50 ranking analysis allowed the selection of the candidate octapeptide containing two (S) 22-amino-4-pentynoic acid (L-Pra) residues Ac-PraRRN(Glc) GHT-Pra-NH2, with an IC50 in the nanomolar range. This peptide was adequately modified for solidphase ELISA (SP-ELISA) and surface plasmon resonance (SPR) experiments. Pra-RRN(Glc) GHT-Pra-NH2 peptide was modified with an alkyl chain linked to the N-terminus, favoring immobilization on solid phase in SP-ELISA and differentiating IgG antibody recognition between patients and healthy blood donors with a high specificity. However, this peptide displayed a loss in IgM specificity and sensitivity. Moreover, an analog was obtained after modification of the octapeptide candidate Ac-Pra-RRN(Glc) GHT-Pra-NH2 to favor immobilization on SPR sensor chips. SPR technology allowed us to determine its affinity (K-D=16.4 nM), 2.3 times lower than the affinity of the original glucopeptide CSF114(Glc) (K-D=7.1 nM). (C) 2015 Wiley Periodicals, Inc.
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