title: Serum complement enhances the responses of genotoxin- and oxidative stress-sensitive Escherichia coli bioreporters.
abstract: Bacterial bioreporters are limited in their abilities to detect large polar molecules due to their membrane selectivity. In this study, the activity of serum complement was used to bypass this undesired selectivity. Initially, the serum complement activity was assessed using the responses of a bacterial bioreporter harboring a recA::luxCDABE transcriptional fusion when exposed to the chemotherapy drug, mitomycin C (MMC). Using 50 °C-treated serum, the limit of detection for this bacterial sensor was lowered by nearly 450-fold, from 31 μg/L to 0.07 μg/L MMC. Real-time quantitative PCR demonstrated that serum-treated cultures responded more strongly to 100 μg/L MMC, with 3.1-fold higher recA expression levels. Subsequent experiments with other bioreporter strains also found enhanced sensitivities and responses. Finally, combining each of the above findings, tests were performed to demonstrate the potential application of the recA::luxCDABE bioreporter within a lab-on-a-CD platform as a point-of-care diagnostic to measure chemotherapeutic drug concentrations within blood.
authors.affiliation: School of Nano-Bioscience and Chemical Engineering, Ulsan National Institute of Science and Technology, Banyeon-ri 100, Ulsan 689-798, Republic of Korea.
authors.forename: Siseon
authors.initials: S
authors.lastname: Lee
authors.forename: Mary
authors.initials: M
authors.lastname: Amasia
authors.forename: Marc
authors.initials: M
authors.lastname: Madou
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Mitchell
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000032:analysis* / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D003165:Complement System Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D057230:Limit of Detection; D044085:Microfluidics / Q000379:methods; D016685:Mitomycin / Q000032:analysis* / Q000378:metabolism; D009153:Mutagens / Q000032:analysis* / Q000378:metabolism; D018384:Oxidative Stress; D010539:Permeability; D011938:Rec A Recombinases / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000903:Antibiotics, Antineoplastic; D029968:Escherichia coli Proteins; D009153:Mutagens; D011993:Recombinant Fusion Proteins; D016685:Mitomycin; D003165:Complement System Proteins; D011938:Rec A Recombinases