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Merck
CN

Rapid quantification of prion proteins using resistive pulse sensing.

The Analyst (2020-02-18)
Matthew J Healey, Muttuswamy Sivakumaran, Mark Platt
ABSTRACT

Prion diseases are a group of fatal transmissible neurological conditions caused by the change in conformation of intrinsic cellular prion protein (PrPC). We present a rapid assay using aptamers and resistive pulse sensing, RPS, to extract and quantify PrPC from complex sample matrices. We functionalise the surface of superparamagnetic beads, SPBs, with a DNA aptamer. First SPB's termed P-beads, are used to pre-concentrate the analyte from a large sample volume. The PrPC protein is then eluted from the P-beads before aptamer modified sensing beads, S-beads, are added. The velocity of the S-beads through the nanopore reveals the concentration of the PrPC protein. The process is done in under an hour and allows the detection of picomol's of protein.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
γ-Globulins from human blood, ≥99% (electrophoresis)
Sigma-Aldrich
Albumin from human serum, lyophilized powder, ≥97% (agarose gel electrophoresis)
Sigma-Aldrich
Fibrinogen from human plasma, 50-70% protein (≥80% of protein is clottable)
Sigma-Aldrich
Phosphate buffered saline, tablet
Sigma-Aldrich
Triton X-100, laboratory grade