2020
DOI: 10.1016/j.snb.2019.127228
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Identification of protein-protein interactions using a magnetic modulation biosensing system

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Cited by 13 publications
(23 citation statements)
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“…PfiA and other antitoxins of type II TA systems are proteins that inhibit their cognate toxin activity by binding to it and creating a protein–protein complex (Unterholzner et al ., 2013b). A magnetic modulation biosensing (MMB) system (Danielli et al ., 2010; Verbarg et al ., 2017; Roth et al ., 2020) was used to determine whether PfiT and PfiA bind to each other. His‐tagged PfiT was co‐overexpressed with Flag‐tagged PfiA.…”
Section: Resultsmentioning
confidence: 99%
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“…PfiA and other antitoxins of type II TA systems are proteins that inhibit their cognate toxin activity by binding to it and creating a protein–protein complex (Unterholzner et al ., 2013b). A magnetic modulation biosensing (MMB) system (Danielli et al ., 2010; Verbarg et al ., 2017; Roth et al ., 2020) was used to determine whether PfiT and PfiA bind to each other. His‐tagged PfiT was co‐overexpressed with Flag‐tagged PfiA.…”
Section: Resultsmentioning
confidence: 99%
“…Bacteria were then diluted 1:100 into M9 with l ‐(+)‐arabinose (33.3 μM) and grown to a turbidity of 0.5 at 595 nm. 1.5 OD (at 595 nm) of bacteria was taken from each strain and proteins were then extracted as previously described (Roth et al ., 2020).…”
Section: Methodsmentioning
confidence: 99%
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“…Then, the complexes were incubated for 20 min at room temperature with 100 µL of 1 µg/mL of streptavidin R-phycoerythrin (SA-PE, PJRS20-1, Agilent Technologies, Santa Clara, CA, USA). Subsequently, after a single buffer replacement to remove unbound fluorescent molecules, 100 µL of the final solution was loaded into a borosilicate glass cuvette and measured in the MMB system [ 25 ]. The LoD was calculated as three standard deviations over the blank measurement.…”
Section: Methodsmentioning
confidence: 99%
“…Previously, we introduced a novel platform that uses magnetically modulated biosensors (MMB) to rapidly detect very low concentrations of antibodies [ 20 ], proteins [ 21 ], and specific nucleic acid sequences [ 22 , 23 , 24 ]. We have shown that the MMB can rapidly identify protein-protein interactions (PPI) in vitro with very high sensitivity [ 25 ]. Here, we demonstrate for the first time the applicability of the biosensors for rapid and highly sensitive screening of PPI inhibitors.…”
Section: Introductionmentioning
confidence: 99%