The Diversity of a Polyclonal FluCell-SELEX Library Outperforms Individual Aptamers as Emerging Diagnostic Tools for the Identification of Carbapenem Resistant Pseudomonas aeruginosa.
Dennis KubiczekHeinz RaberNicholas BodenbergerThomas OswaldMelis SahanDaniel MayerSebastian WieseSteffen StengerTanja WeilFrank RosenauPublished in: Chemistry (Weinheim an der Bergstrasse, Germany) (2020)
Textbook procedures require the use of individual aptamers enriched in SELEX libraries which are subsequently chemically synthesized after their biochemical characterization. Here we show that this reduction of the available sequence space of large libraries and thus the diversity of binding molecules reduces the labelling efficiency and fidelity of selected single aptamers towards different strains of the human pathogen Pseudomonas aeruginosa compared to a polyclonal aptamer library enriched by a whole-cell-SELEX involving fluorescent aptamers. The library outperformed single aptamers in reliable and specific targeting of different clinically relevant strains, allowed to inhibit virulence associated cellular functions and identification of bound cell surface targets by aptamer based affinity purification and mass spectrometry. The stunning ease of this FluCell-SELEX and the convincing performance of the P. aeruginosa specific library may pave the way towards generally new and efficient diagnostic techniques based on polyclonal aptamer libraries not only in clinical microbiology.
Keyphrases
- pseudomonas aeruginosa
- nucleic acid
- escherichia coli
- gold nanoparticles
- cell surface
- cystic fibrosis
- mass spectrometry
- biofilm formation
- sensitive detection
- label free
- endothelial cells
- acinetobacter baumannii
- magnetic nanoparticles
- single cell
- staphylococcus aureus
- high resolution
- cell therapy
- stem cells
- cancer therapy
- living cells
- high performance liquid chromatography
- induced pluripotent stem cells
- capillary electrophoresis
- transcription factor
- gas chromatography
- amino acid
- dna binding
- binding protein
- fluorescent probe