Breakthrough microfluidics-based high-performance liquid chromatography/mass spectrometry technology is expected to significantly increase the speed, ease and productivity of proteomics research.
Agilent Technologies has introduced the first high-performance liquid chromatography (HPLC)-chip/mass spectrometry (MS) system for protein identification.
This 'breakthrough' microfluidics-based LC/MS technology is expected to significantly increase the speed, ease and productivity of proteomics research.
Many biomedical researchers employ proteomics, the systematic analysis of protein expression, structure and function, to study the underlying causes of diseases such as cancer.
Traditional techniques used to prepare protein samples for MS analysis, such as nanoflow LC, require complex arrangements of equipment in order to improve separation power, and consume relatively large quantities of sample.
Agilent's innovative new system simplifies LC/MS, heightens separation power and sensitivity, and reduces required sample sizes by integrating multiple functionalities on a single microfluidics-based chip.
"The compact footprint of the Agilent microfluidic-LC/MS system, combined with its sensitivity, modularity and ease of operation, provides significant advantages for the analysis of complex protein digests and represents a leading-edge technology in proteomics discovery programmes," said Professor Pierre Thibault of the Institute for Research in Immunology and Cancer at the University of Montreal.
The HPLC chip for protein identification.
Agilent's Protein ID HPLC-Chip (G4240A) offers greater peptide coverage and higher protein identification compared with nanoflow LC/MS.
Smaller than a credit card, the company's unique HPLC-Chip for protein identification seamlessly integrates sample enrichment and separation capability of a nanoflow LC system with the intricate connections and spray tips used in electrospray mass spectrometry.
The technology is said to eliminate 50% of the fittings and connections typically required in an LC/MS system, dramatically reducing the possibility of leaks and dead volumes, significantly simplifying workflow, and increasing sensitivity and reliability during analysis.
To enable compliance and experiment management, the HPLC-Chip includes a radio frequency tag that contains information such as chip ID and type, documentation, diagnostic and operational data, and additional memory for user comments.
The RF tag is read each time the chip is inserted into the HPLC-Chip Cube, and information is updated on the tag before chip removal.
The Protein ID HPLC-Chip has a 43mmx75um column packed with 5um Zorbax Stable Bond 300A C-18 reverse phase material.
The HPLC-Chip Cube.
The second component of the HPLC-Chip/MS system is the HPLC-Chip/MS Cube interface, which is mounted on Agilent's Nanoflow Proteomics Solution XCT+ ion trap mass spectrometer.
In addition to positioning the sprayer tip orthogonal to the MS inlet to achieve maximum sensitivity and robustness, the HPLC-Chip Cube MS interface automatically loads and seals the chip, establishing high-pressure, leak-tight fluid connections.
HPLC-Chip replacement is simple and can be completed in a few seconds, as opposed to much longer times required to change out LC columns, says Agilent.
The HPLC-Chip Cube is currently available as a standard module within the Agilent 1100 series LC System for MS.
"The HPLC-Chip helps meet the ever-increasing productivity demands of our customers," said Georges Gauthier, HPLC-Chip technology product manager at Agilent.
"The first HPLC-Chip available is for proteomics applications, but the technology has potential uses across a wide range of other applications in which LC/MS is used, including pharmaceutical development and manufacturing, combinatorial chemistry, compound analysis, food safety, environmental monitoring and homeland security."
