Selection of aptamers against live bacterial cells

Camille L.A. Hamula, Hongquan Zhang, Le Luo Guan, Xing Fang Li, X. Chris Le

Research output: Contribution to journalArticlepeer-review

130 Scopus citations

Abstract

Single-stranded DNA or RNA aptamer molecules have usually been selected against purified target molecules. To eliminate the need of purifying target molecules on the cell surface, we have developed a selection technique using live bacterial cells in suspension as targets, to select for ssDNA aptamers specific to cell surface molecules. Lactobacillus acidophilus cells were chosen to demonstrate proof of principle based on their high abundance of surface molecules (potential targets). Aptamer pools obtained after 6-8 rounds of selection demonstrated high affinity for and selective binding with L. acidophilus cells when tested via flow cytometry, microscopy, and fluorescence measurements. Out of 27 aptamers that were cloned and sequenced, one sequence, hemag1P, was found to bind to L. acidophilus much more strongly and specifically than other cells tested. This aptamer was predicted to have a tight hairpin secondary structure. On average, an estimated 164 ± 47 aptamer molecules were bound to a target cell with an apparent Kd of 13 ± 3 nM. A likely putative molecular target of hemag1P is the S-layer protein on the cell surface.

Original languageEnglish
Pages (from-to)7812-7819
Number of pages8
JournalAnalytical Chemistry
Volume80
Issue number20
DOIs
StatePublished - 15 Oct 2008
Externally publishedYes

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