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Microfluidic Approaches to Multiplexing Heterogeneous Protein Assays

  • Author(s): Gerver, Rachel Ellen
  • Advisor(s): Herr, Amy E
  • et al.
Abstract

While genetic sequencing and other technologies have enabled the investigation of DNA and RNA at an unprecedented scale, new tools are needed to more rapidly advance proteomics research. New protein detection technologies can enable improved disease biomarker identification, more efficient antibody epitope mapping for vaccine development, better point of care diagnostic assays, and accelerated drug discovery. Microfluidic approaches to protein measurements can offer advantages over benchtop methods in terms of faster assay times, increased multiplexing, reduced consumption of sample and reagents, compatibility with full automation, and potential for implementation even in locations with limited lab infrastructure.

This dissertation presents the development of three novel microfluidic platforms for multiplexed protein detection in biological samples:

1) A microfluidic Western blot for low-molecular-mass proteins that enables the separation of proteins down to 6.5kDa with 40% higher separation resolution and a >100-fold improved signal to noise ratio in small-pore-size gels compared to previous approaches.

2) A microfluidic device for synthesizing spectrally encoded microspheres for large scale parallel peptide synthesis and other biological multiplexing applications with the potential to code up to millions of uniquely identifiable microspheres through the use of lanthanide nanoparticles

3) Initial development towards a microfluidic tool for PSA glycan specific isoform identification for improved prostate cancer diagnosis and prognosis.

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