Expanding the limits of routine proteomics using 50 cm micro Pillar Array Column (µPAC™)
The practice of bottom-up proteomics relies to a large extent on the separation performance that can be achieved with state-of-the art nano LC-MS/MS equipment. Depending on the sample complexity or the instrument time that can be dedicated to a certain sample, different LC columns and corresponding LC-MS/MS methods are often required. When aiming for comprehensive proteome analysis with deep coverage, relatively long columns (lengths up to 75 cm) are typically operated with long and shallow solvent gradients, delivering the highest chromatographic performance. However, daily routine proteome analysis often deals with much less complex samples or demands increased sample throughput, making total analysis times above 120 min undesirable or even impossible.
Complementary to its landmark 200 cm long column which is ideally suited to perform comprehensive and sometimes time consuming proteome research, PharmaFluidics also offers a 50 cm long μPAC™ column which can be used in a more routine research setting. With an internal volume of 3 μL, this column is perfectly suited to perform high throughput analyses with shorter gradient solvent times (30, 60 and 90 minute gradients) and it can be used over a wide range of flow rates, between 100 and 2000 nL/min.
In this webinar, Dr. Geert van Raemdonck and Christof Mitterer of PharmaFluidics explained the principles of the 50 cm µPAC™ column and presented routine proteomics data obtained on complex biological samples using micro-chip based pillar array columns.