Development of electrochemical methods to enzymatically detect traumatic brain injury biomarkers

Abstract

As awareness of traumatic brain injury (TBI) increases, the need to detect mild forms and distinguish between the different severities of TBI becomes more apparent. The goal of this work is to develop a point-of-care sensor to detect whole blood biomarkers for rapid and sensitive diagnosis of TBI severity. Presented herein is the enzymatic detection of norepinephrine through the use of immobilization chemistry and impedance techniques. Sustained elevation of norepinephrine concentrations in the blood has been correlated to negative long-term outcomes in TBI cases, often resulting in permanent cognitive or physical deficits.Novel analysis techniques have been used to identify an optimal binding frequency (371.1 Hz) of norepinephrine to the immobilized enzyme on a gold disk electrode. This form of analysis yielded a logarithmic fit characterized by exceptional responsivity (20.89 ohms/pgmL -1 ), reproducibility (R 2 = 0.96), and lower limit of detection (98 pg/mL) first in purified samples, then in rabbit whole blood solutions. Once the optimal binding frequency was determined, the preliminary use of an impedance-time technique was attempted in this work. This technique more closely resembles the amperometric detection method used in commercial self-monitoring blood glucose meters, allowing for continuous or instantaneous measurement of blood borne biomarkers without compromising sensitivity. Future directions include exploration of simultaneous multi-marker detection with the impedance-time technique and experimentation with novel mesoporous materials to filter large blood components.

Current and Future Applications of Biomedical Engineering for Proteomic Profiling: Predictive Biomarkers in Neuro-Traumatology

“…The 16 articles selected fall into two main categories: 8 clinical papers and 8 laboratory studies on animal models of TBI [ 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 ]. Briefly, the 8 clinical studies identified have described four main technologies for identification of predictive biomarkers: (a) Microvescicle/Exosome, (b) MicroRNA, (c) Mass Spectroscopy, and (d) Multiplexing and Immunoassays [ 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 ].…”

“…Briefly, the 8 clinical studies identified have described four main technologies for identification of predictive biomarkers: (a) Microvescicle/Exosome, (b) MicroRNA, (c) Mass Spectroscopy, and (d) Multiplexing and Immunoassays [ 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 ]. On the other hand, the methodology of the 8 laboratory studies was based on (a) MicroRNA, (b) Mass Spectroscopy, and (c) Multiplexing and Immunoassays; those latter investigated several different animal models of TBI ranging from mild to severe TBI including blunt traumas and diffuse axonal injuries; a wide range of animals were investigated in those studies, they included small size animals such as mice and rats but also rabbits and larger size animals such as pigs [ 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 ].…”

“…So far, an heterogeneous group of polypeptides emerged as promising biomarkers for TBIs of different severities; this group includes structural proteins and enzymes reflecting either glial (S-100β, Tau proteins, GFAP, but also Rho-associated protein kinase 2 and carbonic anhydrase-I) or neuronal (peroxiredoxin-2, synaptosomal-associated protein 25, and microtubule-associated protein 1B) wellbeing or damage, depending on their variation from physiological range [ 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 41 , 42 , 43 , 44 , 45 ]. Since many neurological and behavioral abnormalities observed in patients with TBI may in fact be transient or permanent, and are the visible consequences of the cellular, sub-cellular, and molecular pathological processes and their evolution with time, investigations of animal models still result emphasize the paramount importance of approaching all of them in a more comprehensive way.…”

Feasibility of progesterone treatment for ischaemic stroke

“…The ability to detect efficacy following any intervention probably requires assessment of relevant biomarkers that would clearly assist diagnosis and outcome assessment. Although potential biomarkers have been identified in experimental and clinical TBI studies, 14 , 15 they are still lacking appropriate validation and implementation. 16 Ideally, clinical studies would contain a variety of measures in order to better characterize TBI and facilitate individualized treatment.…”

Electrochemical-Nucleic Acid Detection with Enhanced Specificity and Sensitivity

“…The electrochemical technique EIS is used as a potential method of detection that offers DNA hybridization sensing up to lower limit of detection 10 nM concentration [9].…”