title: A fully-automated, six-plex single molecule immunoassay for measuring cytokines in blood.
abstract: We report a system and assay for performing fully-automated measurement of 6 proteins simultaneously with single molecule sensitivity. The system combines handling of samples, reagents, and consumables, with a module for imaging single molecule arrays (Simoa) to enable immunoassays that have high sensitivity (~fg/mL), are multiplexed, and are fully-automated. A 6-plex cytokine Simoa assay for IL-6, TNF-α, GM-CSF, IL-10, IL-1β, and IL-1α was developed on the system. The assays had limits of detection in the range 0.01-0.03pg/mL, and the average imprecision (CV) of the Simoa signal was 4.2%. This assay was used to measure the concentrations of these cytokines in the plasma of patients with Crohn's Disease (CD), before and after treatment with anti-TNF-α antibody drugs, and in the serum of Type 1 diabetics. Concentrations of TNF-α and IL-6 in the CD samples determined using the fully-automated, multiplex Simoa assay had good correlation with the manual, single-plex assays previously reported. Drug treatment caused reductions in the mean concentration of all 6 cytokines in the plasma of CD patients. The concentrations of 4 cytokines were significantly higher in diabetics compared to healthy controls. The system could enable the widespread, multiplexed measurement of protein biomarkers with low abundance.
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Rivnak
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Rissin
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Cheuk W
authors.initials: CW
authors.lastname: Kan
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Linan
authors.initials: L
authors.lastname: Song
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Fishburn
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Piech
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Todd G
authors.initials: TG
authors.lastname: Campbell
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Derek R
authors.initials: DR
authors.lastname: DuPont
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Melissa
authors.initials: M
authors.lastname: Gardel
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Sean
authors.initials: S
authors.lastname: Sullivan
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Brian A
authors.initials: BA
authors.lastname: Pink
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: Carlos G
authors.initials: CG
authors.lastname: Cabrera
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: David R
authors.initials: DR
authors.lastname: Fournier
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA. Electronic address: dduffy@quanterix.com.
authors.forename: David C
authors.initials: DC
authors.lastname: Duffy
mesh_terms: D001331:Automation; D003424:Crohn Disease / Q000097:blood; D016207:Cytokines / Q000097:blood*; D003920:Diabetes Mellitus / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D057188:Workflow
chemical_list: D016207:Cytokines
keywords: Automation; Digital ELISA; Multiplexed immunoassays; Simoa