title: Recognition of bacterial lipopolysaccharide using bacteriophage-adhesin-coated long-period gratings.
abstract: In this paper we present a new type of highly sensitive label-free sensor based on long-period gratings (LPG) coated with T4 bacteriophage (phage) adhesin. The adhesin (gp37) binds Escherichia coli B (E. coli B) by recognizing its bacterial lipopolysaccharide (LPS). The LPG biofunctionalization methodology is based on coating LPG surface with nickel ions capable of gp37 histidine tag reversible binding. For the first time recombinant adhesive phage protein has been used as a receptor molecule in biosensing scheme. The specificity of LPS binding by adhesin has been tested with LPG-based device and confirmed using Western blot, Enzyme-Linked Immunosorbent Assay (ELISA) and BIACORE methods. The LPG-based sensor can measure bacterial contamination in real time and with a high accuracy. We show that T4 phage adhesin binds E. coli B LPS in its native or denatured form. The binding is highly specific and irreversible. The applied procedure allows for obtaining reusable biosensors.
authors: Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wrocław, Poland. Electronic address: ezuziak@iitd.pan.wroc.pl.
authors: Ewa
authors: E
authors: Brzozowska
authors: Institute of Microelectronics and Optoelectronics, Warsaw University of Technology, Koszykowa 75, 00-662 Warsaw, Poland. Electronic address: msmietan@elka.pw.edu.pl.
authors: Mateusz
authors: M
authors: Śmietana
authors: National Institute of Telecommunications, Szachowa 1, 04-894 Warsaw, Poland; Centre de recherche en photonique, Université du Québec en Outaouais, 101 rue Saint-Jean-Bosco, Gatineau, QC, Canada J8X 3X7.
authors: Marcin
authors: M
authors: Koba
authors: Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wrocław, Poland.
authors: Sabina
authors: S
authors: Górska
authors: Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wrocław, Poland.
authors: Krzysztof
authors: K
authors: Pawlik
authors: Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wrocław, Poland.
authors: Andrzej
authors: A
authors: Gamian
authors: Centre de recherche en photonique, Université du Québec en Outaouais, 101 rue Saint-Jean-Bosco, Gatineau, QC, Canada J8X 3X7.
authors: Wojtek J
authors: WJ
authors: Bock
mesh_terms: D017122:Bacteriophage T4 / Q000737:chemistry*; D015374:Biosensing Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000737:chemistry / Q000302:isolation & purification*; D008070:Lipopolysaccharides / Q000737:chemistry / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D008070:Lipopolysaccharides; D011994:Recombinant Proteins; C036757:lipopolysaccharide B
keywords: Bacterial lipopolysaccharide (LPS); Bacteriophage adhesin; Biosensors; Label-free sensing; Long-period gratings (LPG); Optical fiber sensors