Aggregation and Oligomerization Characterization of ß-Lactoglobulin Protein Using a Solid-State Nanopore Sensor

Mitu C. Acharjee, Brad Ledden, Brian Thomas, Xianglan He, Troy Messina, Jason Giurleo, David Talaga, Jiali Li

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Protein aggregation is linked to many chronic and devastating neurodegenerative human diseases and is strongly associated with aging. This work demonstrates that protein aggregation and oligomerization can be evaluated by a solid-state nanopore method at the single molecule level. A silicon nitride nanopore sensor was used to characterize both the amyloidogenic and native-state oligomerization of a model protein ß-lactoglobulin variant A (βLGa). The findings from the nanopore measurements are validated against atomic force microscopy (AFM) and dynamic light scattering (DLS) data, comparing βLGa aggregation from the same samples at various stages. By calibrating with linear and circular dsDNA, this study estimates the amyloid fibrils’ length and diameter, the quantity of the βLGa aggregates, and their distribution. The nanopore results align with the DLS and AFM data and offer additional insight at the level of individual protein molecular assemblies. As a further demonstration of the nanopore technique, βLGa self-association and aggregation at pH 4.6 as a function of temperature were measured at high (2 M KCl) and low (0.1 M KCl) ionic strength. This research highlights the advantages and limitations of using solid-state nanopore methods for analyzing protein aggregation.

Original languageEnglish
Article number81
JournalSensors (Switzerland)
Volume24
Issue number1
DOIs
StatePublished - Jan 2024

Keywords

  • protein aggregation
  • protein oligomerization
  • protein volume
  • solid-state nanopore
  • ß-lactoglobulin

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