2. Academic Validation
  3. A fluorescence sandwich immunoassay for the real-time continuous detection of glucose and insulin in live animals

A fluorescence sandwich immunoassay for the real-time continuous detection of glucose and insulin in live animals

  • Nat Biomed Eng. 2021 Jan;5(1):53-63. doi: 10.1038/s41551-020-00661-1.
Mahla Poudineh  # 1,  Caitlin L Maikawa  # 2,  Eric Yue Ma 1,  Jing Pan 1,  Dan Mamerow 3,  Yan Hang 4,  Sam W Baker 5,  Ahmad Beirami 6,  Alex Yoshikawa 7,  Michael Eisenstein 1 8,  Seung Kim 4,  Jelena Vučković 1 9,  Eric A Appel 10 11 12,  H Tom Soh 13 14 15
Affiliations

Affiliations

  • 1 Department of Electrical Engineering, Stanford University, Stanford, CA, USA.
  • 2 Department of Bioengineering, Stanford University, Stanford, CA, USA.
  • 3 Department of Chemical Engineering, University of California, Santa Barbara, Santa Barbara, CA, USA.
  • 4 Department of Developmental Biology, Stanford University, Stanford, CA, USA.
  • 5 Department of Comparative Medicine, Stanford University, Stanford, CA, USA.
  • 6 Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA, USA.
  • 7 Department of Chemical Engineering, Stanford University, Stanford, CA, USA.
  • 8 Department of Pediatrics (Endocrinology), Stanford University, Stanford, CA, USA.
  • 9 Ginzton Lab, Stanford University, Stanford, CA, USA.
  • 10 Department of Bioengineering, Stanford University, Stanford, CA, USA. [email protected].
  • 11 Department of Pediatrics (Endocrinology), Stanford University, Stanford, CA, USA. [email protected].
  • 12 Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA. [email protected].
  • 13 Department of Electrical Engineering, Stanford University, Stanford, CA, USA. [email protected].
  • 14 Department of Radiology, Stanford University, Stanford, CA, USA. [email protected].
  • 15 Chan Zuckerberg Biohub, San Francisco, CA, USA. [email protected].
  • # Contributed equally.
PMID: 33349659 DOI: 10.1038/s41551-020-00661-1
Abstract

Biosensors that continuously measure circulating biomolecules in real time could provide insights into the health status of patients and their response to therapeutics. But biosensors for the continuous real-time monitoring of analytes in vivo have only reached nanomolar sensitivity and can measure only a handful of molecules, such as glucose and blood oxygen. Here we show that multiple analytes can be continuously and simultaneously measured with picomolar sensitivity and sub-second resolution via the integration of Aptamers and Antibodies into a bead-based fluorescence sandwich immunoassay implemented in a custom microfluidic chip. After an incubation time of 30 s, bead fluorescence is measured using a high-speed camera under spatially multiplexed two-colour laser illumination. We used the assay for continuous quantification of glucose and Insulin concentrations in the blood of live diabetic rats to resolve inter-animal differences in the pharmacokinetic response to Insulin as well as discriminate pharmacokinetic profiles from different Insulin formulations. The assay can be readily modified to continuously and simultaneously measure other blood analytes in vivo.

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