title: Point-of-care diagnostics for noncommunicable diseases using synthetic urinary biomarkers and paper microfluidics.
abstract: With noncommunicable diseases (NCDs) now constituting the majority of global mortality, there is a growing need for low-cost, noninvasive methods to diagnose and treat this class of diseases, especially in resource-limited settings. Molecular biomarkers combined with low-cost point-of-care assays constitute a potential solution for diagnosing NCDs, but the dearth of naturally occurring, predictive markers limits this approach. Here, we describe the design of exogenous agents that serve as synthetic biomarkers for NCDs by producing urinary signals that can be quantified by a companion paper test. These synthetic biomarkers are composed of nanoparticles conjugated to ligand-encoded reporters via protease-sensitive peptide substrates. Upon delivery, the nanoparticles passively target diseased sites, such as solid tumors or blood clots, where up-regulated proteases cleave the peptide substrates and release reporters that are cleared into urine. The reporters are engineered for detection by sandwich immunoassays, and we demonstrate their quantification directly from unmodified urine; furthermore, capture antibody specificity allows the probes to be multiplexed in vivo and quantified simultaneously by ELISA or paper lateral flow assay (LFA). We tailor synthetic biomarkers specific to colorectal cancer, a representative solid tumor, and thrombosis, a common cardiovascular disorder, and demonstrate urinary detection of these diseases in mouse models by paper diagnostic. Together, the LFA and injectable synthetic biomarkers, which could be tailored for multiple diseases, form a generalized diagnostic platform for NCDs that can be applied in almost any setting without expensive equipment or trained medical personnel.
authors.affiliation: Harvard-Massachusetts Institute of Technology Division of Health Sciences and Technology, Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA 02139.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Warren
authors.forename: Gabriel A
authors.initials: GA
authors.lastname: Kwong
authors.forename: David K
authors.initials: DK
authors.lastname: Wood
authors.forename: Kevin Y
authors.initials: KY
authors.lastname: Lin
authors.forename: Sangeeta N
authors.initials: SN
authors.lastname: Bhatia
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000652:urine*; D015179:Colorectal Neoplasms / Q000175:diagnosis* / Q000652:urine; D019937:Diagnostic Techniques and Procedures; D006801:Humans; D007118:Immunoassay; D051379:Mice; D044085:Microfluidics / Q000379:methods*; D053758:Nanoparticles / Q000008:administration & dosage; D010455:Peptides / Q000378:metabolism; D019095:Point-of-Care Systems; D012372:ROC Curve; D018709:Statistics, Nonparametric; D013927:Thrombosis / Q000175:diagnosis* / Q000652:urine
chemical_list: D015415:Biomarkers; D010455:Peptides
keywords: engineered disease reporter; global health; image-free diagnostic; protease nanosensor; urine biomarker