1. Development of a Microsphere-Based System to Facilitate Real-Time Insulin Monitoring.
- Author
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Kahanovitz, Lindy, Seker, Erkin, Marks, Robert S, Yarmush, Martin L, Konry, Tania, and Russell, Steven J
- Subjects
Humans ,Insulin ,Hypoglycemic Agents ,Microfluidic Analytical Techniques ,Microspheres ,Lab-On-A-Chip Devices ,insulin ,insulin analogs ,microfluidic ,microsphere ,pharmacokinetics ,Biotechnology ,Bioengineering ,Diabetes ,Nutrition and Dietetics - Abstract
BackgroundThe speed of insulin absorption after subcutaneous delivery is highly variable. Incorrect assumptions about insulin pharmacokinetics compromise effective glycemic regulation. Our ultimate goal is to develop a system to monitor insulin levels in vivo continuously, allowing pharmacokinetic parameters to be calculated in real time. We hypothesize that a bead-based detection system can be run on a flow-through microfluidic platform to measure insulin in subcutaneous fluid sampled via microdialysis. As a first step in development, we focused on microsphere-based measurement of insulin.MethodsPolystyrene microspheres coated with an anti-insulin monoclonal antibody were exposed to insulin-containing solutions, and after addition of a fluorescently labeled anti-insulin monoclonal antibody with a distinct epitope, bead-associated fluorescence was detected by fluorescence microscopy in 96-well plates or in a flow-through, microfluidic platform.ResultsThe bead detection system in plates had a linear range in buffer for regular human insulin (RHI), insulin lispro, and insulin aspart of 15-1115 µIU/ml, 14-976 µIU/ml, and 25-836 µIU/ml, respectively. Measurement on plasma samples demonstrated proportionality between basal and peak insulin levels similar to the laboratory reference method. Preliminary results in a polydimethylsiloxane-based, flow-through, microfluidic platform showed a strong signal at peak insulin levels.ConclusionsWe have developed a microsphere-based system to rapidly measure levels of insulin and insulin analogs. We have further demonstrated proof of concept that this bead detection system can be implemented in a lab-on-a-chip format, which will be further developed and combined with microdialysis for real-time monitoring of insulin in vivo.
- Published
- 2016