title: In vitro evaluation of fluorescence glucose biosensor response.
abstract: Rapid, accurate, and minimally-invasive glucose biosensors based on Förster Resonance Energy Transfer (FRET) for glucose measurement have the potential to enhance diabetes control. However, a standard set of in vitro approaches for evaluating optical glucose biosensor response under controlled conditions would facilitate technological innovation and clinical translation. Towards this end, we have identified key characteristics and response test methods, fabricated FRET-based glucose biosensors, and characterized biosensor performance using these test methods. The biosensors were based on competitive binding between dextran and glucose to concanavalin A and incorporated long-wavelength fluorescence dye pairs. Testing characteristics included spectral response, linearity, sensitivity, limit of detection, kinetic response, reversibility, stability, precision, and accuracy. The biosensor demonstrated a fluorescence change of 45% in the presence of 400 mg/dL glucose, a mean absolute relative difference of less than 11%, a limit of detection of 25 mg/dL, a response time of 15 min, and a decay in fluorescence intensity of 72% over 30 days. The battery of tests presented here for objective, quantitative in vitro evaluation of FRET glucose biosensors performance have the potential to form the basis of future consensus standards. By implementing these test methods for a long-visible-wavelength biosensor, we were able to demonstrate strengths and weaknesses with a new level of thoroughness and rigor.
authors.affiliation: Center for Devices and Radiological Health, Food and Drug Administration, Silver Spring, MD 20993, USA. Mamdouh.Aloraefy@fda.hhs.gov.
authors.forename: Mamdouh
authors.initials: M
authors.lastname: Aloraefy
authors.affiliation: Center for Devices and Radiological Health, Food and Drug Administration, Silver Spring, MD 20993, USA. Joshua.Pfefer@fda.hhs.gov.
authors.forename: T Joshua
authors.initials: TJ
authors.lastname: Pfefer
authors.affiliation: Department of Biomedical Engineering and Herbert Wertheim College of Medicine, Florida International University, Miami, FL 33174, USA. jramella@fiu.edu.
authors.forename: Jessica C
authors.initials: JC
authors.lastname: Ramella-Roman
authors.affiliation: Center for Devices and Radiological Health, Food and Drug Administration, Silver Spring, MD 20993, USA. Kim.Sapsford@fda.hhs.gov.
authors.forename: Kim E
authors.initials: KE
authors.lastname: Sapsford
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D003208:Concanavalin A / Q000737:chemistry*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation*; D005947:Glucose / Q000032:analysis* / Q000737:chemistry; D066298:In Vitro Techniques; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D003208:Concanavalin A; D005947:Glucose