Eksigent Column Chips for Nano and Micro LC Proteomics Analysis

AB SCIEX has announced the extension of its Eksigent line of column chips for the cHiPLC®-nanoflex system. Five new chips were introduced expanding the flexibility and application of the system for high-performance proteomics analysis including biomarker discovery, validation and verification. The extension adds higher flow micro cHiPLC columns for peptide separation at increased throughput levels, as well as new phases for the analysis of intact proteins, phosphopeptides and glycans. All cHiPLC-nanoflex columns are designed and microfabricated to be robust and highly reproducible, providing high quality separations that are easy to obtain by users at all experience levels.

The first two new chips are nano LC columns – C4 for the analysis of intact proteins and graphitized carbon for phosphopeptides and glycans.

AB SCIEX also previewed three additional new chips at HUPO including 200 µm i.d. C18-CL micro cHiPLC columns in 15 cm as well as a new shorter 5 cm length. These chips are run at 1.0 to 3.5 µL per-minute flow rates, reducing gradient delay times compared to nano flow HPLC, providing higher sample throughput and improved chromatography. The new micro cHiPLC columns deliver robust performance as well as increased speed and throughput for applications such as quantitative biomarker validation.

Trap columns are available for all phases, including the original C18 and C8 columns for peptide analysis.

Eksigent customer Reid Townsend, professor of medicine, cell biology and physiology at the Washington University School of Medicine, and director of their NIH-NCI proteome characterization center, has tested the new 200 µm chips in his laboratory and reports that loading five times more sample on the new chip with a yeast digest yielded five times more protein identifications resulting from three times the number of distinct peptide identifications, compared to a standard 75 µm i.d. chip.

Nanoflow liquid chromatography coupled with nanoelectrospray MS is the method of choice for protein and peptide analysis. The Eksigent cHiPLC-nanoflex System is a docking station for up to three column chips. Eksigent’s proprietary EksPortTM microconnector system ensures reliable and consistent connections to facilitate column changes.

Chips can be interchanged in seconds, even by inexperienced users, with minimal dead volume to compromise resolution. This enables users to easily switch between workflows. The proprietary microfabrication process used for both the trap chips and analytical column chips delivers excellent column-to-column reproducibility.

The cHiPLC-nanoflex can be added to any of Eksigent’s nanoLC systems and interfaced to AB SCIEX TripleTOF™ 5600 for biomarker discovery and the AB SCIEX QTRAP® 5500 for verification and validation, as well as other mass spectrometers.

“Chromatography on the nano and micro scale has traditionally suffered from reproducibility and robustness limitations, and our cHiPLC columns deliver significant improvements,” commented Remco van Soest, director of Eksigent HPLC product management for AB SCIEX. “This is part of our effort to develop and introduce new cHiPLC columns on an ongoing basis to meet the needs of our proteomics customers.”