Microfluidic device for single step measurement of protein C in plasma samples for sepsis prognosis

Lab Chip. 2022 Jun 28;22(13):2566-2577. doi: 10.1039/d1lc01084k.

Abstract

Protein C is a vitamin K dependant protein in plasma that plays an essential role in regulating the coagulation cascade and inflammatory response. As a result of its importance in these roles, it has been suggested as a biomarker for prognosis of patients affected by sepsis. Sepsis is a dysregulated host response to an infection that is the leading cause of mortality in U.S. hospitals and results in the highest cost of hospitalization. It was found that protein C concentration in non-surviving sepsis patients is significantly lower (1.8 μg mL-1) than in survivors and healthy patients who have a protein C concentration of 3.9-5.9 μg mL-1. Current methods for diagnosing sepsis rely on expensive immunoassays or functional assays that require multiple steps for isolation and activation of protein C. We demonstrate in this paper a low cost, single step assay for detection of protein C in blood plasma. This was done by combining isoelectric gates with barium-immobilized metal affinity trapping. The electric field was optimized for use with immobilized metal affinity using COMSOL simulation. The integrated device was tested with samples containing buffered protein C, protein C in the presence of high concentration bovine serum albumin and alpha 1-proteinase inhibitor, and in blood plasma with spiked protein C. The stability of the measured values was tested by monitoring the intensity of a mixture of protein C with BSA and A1PI every minute to determine that measurement after 40 minutes was optimal. The results showed that the device could be used to distinguish a reduction in protein C from 4.46 μg mL-1 to 1.96 μg mL-1 with greater than 98% confidence in plasma making it suitable for sepsis prognosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers
  • Humans
  • Lab-On-A-Chip Devices
  • Prognosis
  • Protein C*
  • Sepsis* / diagnosis
  • alpha 1-Antitrypsin

Substances

  • Biomarkers
  • Protein C
  • alpha 1-Antitrypsin

Grants and funding