Agilent's new application note describes how its Bravo liquid-handling system enables the automation of a SISCAPA Magnetic Bead Workflow for protein biomarker quantification by mass spectrometry.
The authors of the note describe an automated protocol implementing SISCAPA immunoaffinity enrichment of biomarker peptides prior to quantification by MS.
In the described protocol, magnetic beads coated with anti-peptide antibodies were used to bind specific target peptides from tryptic digests of plasma and serum samples, after which the beads were washed to remove unbound peptides and the bound, purified peptides were eluted in small volumes for injection and analysis in an LC/MS/MS system.
Stable isotope-labelled versions of the same peptides was used as internal standards to enable accurate quantification.
The Agilent Bravo automated protocol for this application enabled SISCAPA processing of 96 samples in less than 30m - less than half the time required by traditional manual methods.