Optimizing Multiplexed Detections of Diabetes Antibodies via Quantitative Microfluidic Droplet Array

Small. 2017 Dec;13(46):10.1002/smll.201702323. doi: 10.1002/smll.201702323. Epub 2017 Oct 9.

Abstract

Sensitive, single volume detections of multiple diabetes antibodies can provide immunoprofiling and early screening of at-risk patients. To advance the state-of-the-art suspension assays for diabetes antibodies, porous hydrogel droplets are leveraged in microfluidic serpentine arrays to enhance reagent transport. This spatially multiplexed assay is applied to the detection of antibodies against insulin, glutamic acid decarboxylase, and insulinoma-associated protein 2. Optimization of assay protocol results in a shortened assay time of 2 h, with better than 20 pg mL Supporting Information detection limits across all three antibodies. Specificity and cross-reactivity tests show negligible background, nonspecific antibody-antigen, and nonspecific antibody-antibody bindings. Multiplexed detections are able to measure within 15% of target concentrations from low to high ranges. The technique enables quantifications of as little as 8000 molecules in each 500 µm droplet in a single volume, multiplexed assay format, a breakthrough necessary for the adoption of diabetes panels for clinical screening and monitoring in the future.

Keywords: antibody; diabetes; droplets; microfluidic; multiplexing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / analysis*
  • Antigens / metabolism
  • Cross Reactions
  • Diabetes Mellitus / immunology*
  • Humans
  • Immunoassay
  • Limit of Detection
  • Microfluidics / methods*
  • Polyethylene Glycols / chemistry
  • Time Factors

Substances

  • Antibodies
  • Antigens
  • poly(ethylene glycol)diacrylate
  • Polyethylene Glycols