Selection of an Aptamer against Mouse GP2 by SELEX
スポンサーリンク
概要
- 論文の詳細を見る
Microfold (M) cells are intestinal epithelial cells specialized for sampling and transport of luminal antigens to gut-associated lymphoid tissue for initiation of both mucosal and systemic immune responses. Therefore, M-cell targeted vaccination has the potential to be a better immunization strategy. Glycoprotein 2 (GP2), an antigen uptake receptor for FimH+ bacteria on M cells, can be a good target for this purpose. Aptamers are oligonucleotides that bind to a variety of target molecules with high specificity and affinity. Together with its low toxic feature, aptamers serves as a tool of molecular-targeted delivery. In this study, we used Systematic Evolution of Ligands by EXponential enrichment (SELEX) to isolate aptamers specific to murine GP2 (mGP2). After ten rounds of SELEX, eleven different aptamer sequences were selected. Among them, the most frequently appeared sequence (~60%) were aptamer NO. 1 (Apt1), and the second most (~7%) were aptamer NO. 5 (Apt5). In vitro binding experiment confirmed that only Apt1 and Apt5 specifically bound to mGP2 among eleven aptamers initially selected. Apt1 showed the strongest affinity with mGP2, with the Kd value of 110±2.6 nM evaluated by BIACORE. Binding assays with mutants of Apt1 suggest that, in addition to the loop structure, the nucleotide sequence, AAAUA, in the loop is important for binding to mGP2. Furthermore, this aptamer was able to bind to mGP2 expressed on the cell surface. These results suggest that this mGP2-specific aptamer could serve as a valuable tool for testing M-cell-targeted vaccine delivery in the murine model system.
- 日本細胞生物学会の論文
日本細胞生物学会 | 論文
- テトラヒメナにおけるDNA-核膜複合体の研究 (細胞核内小器官の生物学)
- 核小体におけるリボゾ-ムRNA合成の制御 (細胞核内小器官の生物学)
- 細胞分裂とその調節-分裂装置をめぐって (細胞増殖と分化)
- 細胞雑種研究の現状 (細胞融合)
- 浮遊増殖性癌細胞の無血清培養と培地添加アルブミンの役割 (細胞融合)