Selection of an aptamer against mouse GP2 by SELEX

Misaho Hanazato, Gaku Nakato, Fumiko Nishikawa, Koji Hase, Satoshi Nishikawa, Hiroshi Ohno

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)23-29
Number of pages7
JournalCell structure and function
Volume39
Issue number1
DOIs
Publication statusPublished - 2014 Jan 27
Externally publishedYes

Keywords

  • Aptamer
  • GP2
  • M cells
  • SELEX

ASJC Scopus subject areas

  • Physiology
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Selection of an aptamer against mouse GP2 by SELEX'. Together they form a unique fingerprint.

  • Cite this

    Hanazato, M., Nakato, G., Nishikawa, F., Hase, K., Nishikawa, S., & Ohno, H. (2014). Selection of an aptamer against mouse GP2 by SELEX. Cell structure and function, 39(1), 23-29. https://doi.org/10.1247/csf.13019