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Design and Fabrication of a 3D-Printed Microfluidic Immunoarray for Ultrasensitive Multiplexed Protein Detection.

Keshani Hiniduma1, Ketki S Bhalerao1, Peyahandi I Thilini De Silva1

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Area of Science:

  • Microfluidics and Lab-on-a-Chip Technology
  • Biomarker Detection and Diagnostics
  • 3D Printing and Additive Manufacturing

Background:

  • Microfluidic systems integrate fluid dynamics with multiple scientific disciplines for automated tasks.
  • Low-cost 3D printing significantly enhances the development of microfluidic devices, offering cost, time, and design advantages.
  • 3D-printed microfluidics are particularly beneficial for molecular measurement applications.

Purpose of the Study:

  • To provide a comprehensive tutorial on designing, optimizing, and validating a 3D-printed microfluidic immunoarray.
  • To develop a point-of-care array for the ultrasensitive detection of five protein biomarkers associated with aggressive cancers.
  • To demonstrate the methodology for creating innovative and accurate biomarker analysis tools.

Main Methods:

  • Iterative physical design of the microfluidic array, including microchannels, reagent chambers, and open detection wells with a covering flap.
  • Optimization of signal detection methods to enhance sensitivity and determine the limit of detection (LOD).
  • Generation of calibration plots to assess linear dynamic ranges and validation through spike-recovery studies for accuracy.

Main Results:

  • Optimized design features for uniform signal distribution and assay coverage.
  • Achieved ultrasensitive detection of multiple protein biomarkers with determined LODs and linear dynamic ranges.
  • Spike-recovery studies confirmed the accuracy of the developed immunoarray assay.

Conclusions:

  • The paper successfully showcases the design, optimization, and validation of an innovative 3D-printed microfluidic immunoarray.
  • The optimized parameters, calibration data, and sensitivity metrics provide valuable resources for future biomarker analysis.
  • This work highlights the potential of 3D printing for creating advanced point-of-care diagnostic tools for aggressive cancers.