Speaker: Carol Haney, Agilent Technologies, Inc., 3333 Regency Parkway, Suite 100, Cary, North Carolina 27511

Topic: Fully Automated High Performance (2D) Nano-LC/Ion Trap Mass Spectrometry

Place: Building 549, Auditorium, NCI-Frederick, Frederick, MD

Time: Tuesday, April 9, 2002, at 2:00 PM

Abstract: Nanoscale 2D-LC in combination with nanoelectrospray ion trap MS/MS can be used to identify proteins from complex digest mixtures at low femtomole levels. Skilled operators are typically required to monitor and maintain these powerful systems. Problems such as a partially plugged sprayer needles, passive splitters or tubing may cause unstable flows which result in poor quality electrospray MS/MS results. Passive splitters which use exclusively pressure differences to control nanoflow can not compensate for the above uncontrolled events. The complexity of multiple protein digests requires new technology which includes multi-dimensional chromatographic systems. To get the highest throughput possible the entire system must be automated and robust enough to be operated routinely. A system for a routine environment therefore needs (1) reliable generation of shallow gradients with small delay volumes from 0-60% organic at 100-600 nl/min, (2) direct flow readouts for diagnostics, (3) optimized hardware to maintain trapping / analytical performance, electrospray needles, and a low dispersive system for maximum chromatographic fidelity, and (4) a stable software environment for reliable automation of LC/MS along with fully automatic post-processing protein identifications from digestion mixtures or gel spots.