title: Ultrasensitive microfluidic array for serum pro-inflammatory cytokines and C-reactive protein to assess oral mucositis risk in cancer patients.
abstract: In addition to disease diagnostics, there is a need for biomarkers to predict severity of cancer therapy side effects such as oral mucositis. Oral mucositis is an inflammatory lesion of oral mucosa caused by high-dose chemotherapy and/or radiation that is especially prevalent during oral cancer treatment. We describe here a semi-automated, modular microfluidic immunoarray optimized for ultrasensitive detection of pro-inflammatory cytokines involved in pathobiology of oral mucositis. Our goal is to methodologically identify risk of mucositis early in oral cancer treatment, before the onset of lesions. Biomarkers include tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and C-reactive protein (CRP). Protein analytes were captured from serum in a capture chamber by 1-μm magnetic beads coated with antibodies and enzyme labels. Beads are then transported downstream to a detection chamber containing an eight-sensor array coated with glutathione-coated gold nanoparticles (GSH-AuNP) and a second set of antibodies to capture the beads with analyte proteins. In this first application of the immunoarray to four-protein multiplexed measurements, ultralow detection limits of 10-40 fg mL(-1) in 5 μL serum were achieved for simultaneous detection in 30 min. Mass detection limits were 2.5-10 zmol, as few as 1500 molecules. Accuracy and diagnostic utility of the arrays were demonstrated by correlation of levels of the four biomarker proteins in serum from head and neck cancer patients with results from standard ELISA. This approach may lead to rapid, low-cost estimates of projected risk for severity of oral mucositis in cancer patients to enable improved therapeutic management.
authors.affiliation: Department of Chemistry, University of Connecticut, 55 North Eagleville Road, Storrs, CT, 06269, USA.
authors.forename: Colleen E
authors.initials: CE
authors.lastname: Krause
authors.forename: Brunah A
authors.initials: BA
authors.lastname: Otieno
authors.forename: Gregory W
authors.initials: GW
authors.lastname: Bishop
authors.forename: Gayatri
authors.initials: G
authors.lastname: Phadke
authors.forename: Linda
authors.initials: L
authors.lastname: Choquette
authors.forename: Rajesh V
authors.initials: RV
authors.lastname: Lalla
authors.forename: Douglas E
authors.initials: DE
authors.lastname: Peterson
authors.forename: James F
authors.initials: JF
authors.lastname: Rusling
mesh_terms: D015415:Biomarkers / Q000097:blood; D002097:C-Reactive Protein / Q000032:analysis*; D016207:Cytokines / Q000097:blood*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D018189:Immunomagnetic Separation / Q000295:instrumentation*; D018836:Inflammation Mediators / Q000097:blood; D056656:Lab-On-A-Chip Devices; D040081:Protein Array Analysis / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013280:Stomatitis / Q000097:blood* / Q000175:diagnosis*
chemical_list: D015415:Biomarkers; D016207:Cytokines; D018836:Inflammation Mediators; D002097:C-Reactive Protein