title: Rapid Screening of Antimicrobial Synthetic Peptides.
abstract: Increasing resistance to conventional antibiotics among microorganisms is one of the leading problems of medicine nowadays. Antimicrobial peptides are compounds exhibiting both antibacterial and antifungal activities. However, it is difficult to predict whether a designed new compound would exhibit any biological activity. Moreover, purification of the peptides is one of the most time-consuming and expensive steps of the synthesis that sometimes leads to unnecessary loss of solvents and reagents. In our study we have developed a thin-layer chromatography (TLC) direct bioautography technique for rapid determination of antimicrobial activity of peptides without the necessity of high-performance liquid chromatography purification. In this assay, crude peptides were applied and separated on a TLC plate. Then, pre-prepared plates were dipped into microbial suspension and incubated under optimum conditions for bacteria and fungi as well. The activity of the tested compounds was visualized by spraying the TLC plates with a cell viability reagent, resazurin (7-hydroxy-3H-phenoxazin-3-one 10-oxide). Effectiveness of this assay was compared with minimal inhibitory concentration results obtained by broth microdilution assay. Interestingly, so far such a screening method has not been applied for this group of compounds.
authors.affiliation: Department of Inorganic Chemistry, Faculty of Pharmacy, Medical University of Gdansk, Al. Gen. J. Hallera 107, 80-416 Gdańsk, Poland.
authors.forename: Maciej
authors.initials: M
authors.lastname: Jaskiewicz
authors.affiliation: Department of Inorganic Chemistry, Faculty of Pharmacy, Medical University of Gdansk, Al. Gen. J. Hallera 107, 80-416 Gdańsk, Poland.
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Orlowska
authors.affiliation: Department of Inorganic Chemistry, Faculty of Pharmacy, Medical University of Gdansk, Al. Gen. J. Hallera 107, 80-416 Gdańsk, Poland.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Olizarowicz
authors.affiliation: Department of Inorganic Chemistry, Faculty of Pharmacy, Medical University of Gdansk, Al. Gen. J. Hallera 107, 80-416 Gdańsk, Poland.
authors.forename: Dorian
authors.initials: D
authors.lastname: Migon
authors.affiliation: Lipopharm.pl, ul. Koscielna 16A, 83-210 Zblewo, Poland.
authors.forename: Daria
authors.initials: D
authors.lastname: Grzywacz
authors.affiliation: Department of Inorganic Chemistry, Faculty of Pharmacy, Medical University of Gdansk, Al. Gen. J. Hallera 107, 80-416 Gdańsk, Poland.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Kamysz
keywords: Antimicrobial peptides; Direct bioautography; Peptides; TLC; TLC-DB