title: Detection of the Lipopeptide Pam3CSK4 Using a Hybridized Toll-like Receptor Electrochemical Sensor.
abstract: Electrochemical detection of Pam3CSK4, a synthetic triacylated lipopeptide that mimics the structural moieties of its natural Gram negative bacterial pathogen-associated molecular pattern (PAMP) counterpart, has been achieved using hybridized toll-like receptors (TLR) combining TLR1 and TLR2 onto a single sensor surface. These sensors represent the first hybridized TLR sensors. The limit of detection for Pam3CSK4 attained was 7.5 μg/mL, which is within the same order of magnitude for that of the more labor-intensive and time-consuming cell-assay technique, 2.0 μg/mL. The results gathered in these electrochemical experiments show that sensors fabricated by immobilizing a mixture of cooperative TLR1 and -2 generate higher responses when exposed to the analyte in comparison to the control sensors fabricated using pure TLR1 or -2 standalone. A PAMP selectivity test was carried out in line with our inspiration from the mammalian innate immune response. TLRs1-5 as standalone biorecognition elements and the hybridized "TLR1 and 2" sensor surface were investigated, understanding the known TLR-PAMP interactions, through the exploitation of this electrochemical sensor fabrication technique. The experimental result is consistent with observations from previously published in vivo and in vitro studies, and it is the first demonstration of the simultaneous evaluation of electrochemical responses from multiple, unique fabricated TLR sensor surfaces against the same analyte.
authors.affiliation: Department of Chemistry and Chemical Engineering, Royal Military College of Canada , P.O. Box 17000, Station Forces, Kingston, Ontario K7K 7B4, Canada.|Department of Physical and Environmental Sciences, University of Toronto Scarborough , 1265 Military Trail, Toronto, Ontario M1C 1A4, Canada.
authors.forename: Zhe
authors.initials: Z
authors.lastname: She
authors.affiliation: Department of Chemistry and Chemical Engineering, Royal Military College of Canada , P.O. Box 17000, Station Forces, Kingston, Ontario K7K 7B4, Canada.
authors.forename: Kristin
authors.initials: K
authors.lastname: Topping
authors.affiliation: Department of Physical and Environmental Sciences, University of Toronto Scarborough , 1265 Military Trail, Toronto, Ontario M1C 1A4, Canada.
authors.forename: Tianxiao
authors.initials: T
authors.lastname: Ma
authors.affiliation: Department of Immunology, University of Toronto Scarborough , 1265 Military Trail, Toronto, Ontario M1C 1A4, Canada.
authors.forename: Tiantian
authors.initials: T
authors.lastname: Zhao
authors.affiliation: Department of Physical and Environmental Sciences, University of Toronto Scarborough , 1265 Military Trail, Toronto, Ontario M1C 1A4, Canada.
authors.forename: Wenxia
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Department of Physical and Environmental Sciences, University of Toronto Scarborough , 1265 Military Trail, Toronto, Ontario M1C 1A4, Canada.
authors.forename: Ajar
authors.initials: A
authors.lastname: Kamal
authors.affiliation: Department of Physical and Environmental Sciences, University of Toronto Scarborough , 1265 Military Trail, Toronto, Ontario M1C 1A4, Canada.|Department of Chemistry, University of Toronto , 80 St. George Street, Toronto, M5S 3H6, Canada.
authors.forename: Soha
authors.initials: S
authors.lastname: Ahmadi
authors.affiliation: Department of Chemistry and Chemical Engineering, Royal Military College of Canada , P.O. Box 17000, Station Forces, Kingston, Ontario K7K 7B4, Canada.|Department of Physical and Environmental Sciences, University of Toronto Scarborough , 1265 Military Trail, Toronto, Ontario M1C 1A4, Canada.|Department of Chemistry, University of Toronto , 80 St. George Street, Toronto, M5S 3H6, Canada.
authors.forename: Heinz-Bernhard
authors.identifier: 0000-0002-7149-0110
authors.initials: HB
authors.lastname: Kraatz
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D055664:Electrochemical Techniques / Q000379:methods*; D057230:Limit of Detection; D055666:Lipopeptides / Q000032:analysis* / Q000737:chemistry; D051379:Mice; D051194:Toll-Like Receptor 1 / Q000737:chemistry; D051195:Toll-Like Receptor 2 / Q000737:chemistry
chemical_list: D055666:Lipopeptides; C508075:Pam(3)CSK(4) peptide; D051194:Toll-Like Receptor 1; D051195:Toll-Like Receptor 2