Automation of a Magnetic Bead Workflow for Protein Biomarker Quantification

Agilent Automation Solutions has released a new application note that demonstrates how its Bravo liquid handling system provides an efficient platform for automation of a SISCAPA Magnetic Bead Workflow for protein biomarker quantification by Mass Spectrometry.

Mass Spectrometry (MS) has become the benchmark technology for analysis of peptides based upon its ability to deliver precise quantification, near absolute structural specificity and multiplexing of measurements in a single analytical run. These advantages can be exploited in the analysis of peptides which occur in a single sequence to reveal the concentration of the parent proteins in digests of complex samples such as patient plasma or serum.

The authors of this new application 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 labeled 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 30 minutes, less than half the time required by traditional manual methods.

For a copy of this application note visit www.chem.agilent.com/Library/applications/5990-7360en_lo%20CMS.pdf or contact your local Agilent sales office.