title: One round of SELEX for the generation of DNA aptamers directed against KLK6.
abstract: Kallikrein-related peptidase 6 (KLK6) is an active serine protease that has been implicated in common pathologies, including neurodegenerative disorders such as Parkinson and Alzheimer disease and certain types of cancer. Antibodies, either polyclonal or monoclonal, that exhibit specificity for distinct members of the extended kallikrein family, including KLK6, were developed. With the exception of KLK3/PSA, the identification and generation of aptamers, as potential new tools with improved characteristics demanded for therapeutic and diagnostic applications, has not been explored for KLKs. Here, we report for the first time the identification of novel DNA aptamers against KLK6 that were isolated using a modified systemic evolution of ligands by exponential enrichment technique. The identified aptamers were characterized using fluorescence spectroscopy, competition ELISA, and quartz crystal microbalance, and two aptamers (008 and 022) were found to exhibit high affinity (K(d) in the low nanomolar range) for KLK6. Aptamers were tested for their ability to bind to serum albumin, to demonstrate their specificity for their target, and the possible involvement of such proteins in the transport of aptamers into the bloodstream. The developed aptamers are expected to assist the development of novel diagnostic, biosensing, and therapeutic strategies.
authors.affiliation: Department of Chemistry, The Open University, Milton, Keynes, UK.
authors.forename: Steve
authors.initials: S
authors.lastname: Arnold
authors.forename: Georgios
authors.initials: G
authors.lastname: Pampalakis
authors.forename: Kalliopi
authors.initials: K
authors.lastname: Kantiotou
authors.forename: Dilson
authors.initials: D
authors.lastname: Silva
authors.forename: Celia
authors.initials: C
authors.lastname: Cortez
authors.forename: Sotiris
authors.initials: S
authors.lastname: Missailidis
authors.forename: Georgia
authors.initials: G
authors.lastname: Sotiropoulou
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D015374:Biosensing Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007610:Kallikreins / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008958:Models, Molecular; D011487:Protein Conformation; D052156:SELEX Aptamer Technique / Q000379:methods*; D012709:Serum Albumin / Q000378:metabolism
chemical_list: D052157:Aptamers, Nucleotide; D012709:Serum Albumin; C103551:KLK6 protein, human; D007610:Kallikreins