- Mar 1, 2019

App Note: Nanotrap® Particles for Mass Spec - On-particle digestion

Updated: Nov 6, 2019

APPLICATION NOTE - SKU 33XXX

Nanotrap® Particles for Mass Spectrometry: Performing on-particle protein digestion reduces total processing time and increases unique protein identifications

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Introduction

The Nanotrap® particle technology platform offers a powerful approach for enrichment of low abundance proteins prior to proteolytic digestion, fractionation by liquid chromatography, and analysis by mass spectrometry. For an experimental protocol and results from that protocol, see the October 2018 Ceres Nanosciences application note, “Nanotrap® Particles for Mass Spectrometry: Enrichment of low abundance, low molecular weight proteins from human plasma samples."

In that application note, Nanotrap® particles are incubated with the sample and target proteins are bound to the particles. The Nanotrap® particles are washed and incubated with an elution buffer to release the captured proteins. Then, eluted proteins are subjected to trypsin digestion and desalting. Finally, the digested proteins are analyzed by LC-MS.

This application note presents a simplified “on-particle” digestion method, which reduces processing time by performing protein digestion and clean up while the proteins are still bound to the Nanotrap® particles (Figure 1), and which leads to a larger number of unique proteins identified via LC-MS. This note also compares a 70-minute and 120-minute LC gradient time.