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Open AccessArticle

A Capillary-Perfused, Nanocalorimeter Platform for Thermometric Enzyme-Linked Immunosorbent Assay with Femtomole Sensitivity

1
Department of Biomedical Engineering, Vanderbilt University, Nashville, TN 37235, USA
2
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37235, USA
*
Author to whom correspondence should be addressed.
Biosensors 2020, 10(6), 71; https://doi.org/10.3390/bios10060071
Received: 18 May 2020 / Revised: 18 June 2020 / Accepted: 19 June 2020 / Published: 24 June 2020
Enzyme-catalyzed chemical reactions produce heat. We developed an enclosed, capillary-perfused nanocalorimeter platform for thermometric enzyme-linked immunosorbent assay (TELISA). We used catalase as enzymes to model the thermal characteristics of the micromachined calorimeter. Model-assisted signal analysis was used to calibrate the nanocalorimeter and to determine reagent diffusion, enzyme kinetics, and enzyme concentration. The model-simulated signal closely followed the experimental signal after selecting for the enzyme turnover rate (kcat) and the inactivation factor (InF), using a known label enzyme amount (Ea). Over four discrete runs (n = 4), the minimized model root mean square error (RMSE) returned 1.80 ± 0.54 fmol for the 1.5 fmol experiments, and 1.04 ± 0.37 fmol for the 1 fmol experiments. Determination of enzyme parameters through calibration is a necessary step to track changing enzyme kinetic characteristics and improves on previous methods to determine label enzyme amounts on the calorimeter platform. The results obtained using model-system signal analysis for calibration led to significantly improved nanocalorimeter platform performance. View Full-Text
Keywords: microfabricated calorimeter; ELISA; thermometric ELISA; biosensor; model-assisted signal analysis microfabricated calorimeter; ELISA; thermometric ELISA; biosensor; model-assisted signal analysis
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MDPI and ACS Style

Kazura, E.; Mernaugh, R.; Baudenbacher, F. A Capillary-Perfused, Nanocalorimeter Platform for Thermometric Enzyme-Linked Immunosorbent Assay with Femtomole Sensitivity. Biosensors 2020, 10, 71.

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