Identification of distinct antibody epitopes and mimotopes from a peptide array of 5520 randomly generated sequences

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• 作者：Reineke ; Ulrich ; Ivascu ; Claudia ; Schlief ; Maré ; n ; Landgraf ; Christiane ; Gericke ; Seike ; et. al.
• 关键词：Peptide library ; Peptide array ; SPOT synthesis ; Epitope ; Mimotope ; Antibody ; Molecular recognition ; Molecular diversity ; DMSO ; dimethylsulfoxide ; EDTA ; ethylendiamine-N ; N ; N&prime ; N&prime ; -tetraacetic acid ; HBS ; Hepes (4-(2-hydroxyethly)-1-piperazine etha
• 刊名：Journal of Immunological Methods
• 出版年：2002
• 出版时间：September 1, 2002
• 年：2002
• 卷：267
• 期：1
• 页码：37-51
• 全文大小：988 K

文摘

We used a relatively small library of 5520 randomly generated single 15-mer peptides prepared by SPOT synthesis as an array of 28.5×19.0 cm to identify epitopes for three distinct monoclonal antibodies, namely anti-p24 (human immunodeficiency virus (HIV)-1) monoclonal anibody (mab) CB4-1, anti-interleukin-10 (IL-10) mab CB/RS/13, and anti-transforming growth factor α (TGFα) mab Tab2. Initially identified peptide ligands mostly had very low affinities for the antibodies with dissociation constants around 10⁻⁴ M. Subsequent identification of residues critical for the antibody interactions involved complete L-amino acid substitutional analyses. Several substitutions resulted in analogs with dissociation constants in the low micromolar and high nanomolar range. Specifically binding peptides with key residue patterns matching the wild-type epitopes were identified for all three antibodies. In addition, for antibody CB4-1 mimotopes that showed no homology to the known epitope were selected. Our results suggest that a very limited library diversity, although far from covering the entire sequence repertoire, can suffice to rapidly and economically select peptidic antibody epitopes and mimotopes.