Pooling Spin Column Eluates for Higher Throughput
Perhaps the most efficient method to achieve higher sample throughput is to pool the GPC spin column eluates. Since the anticipated number of hits per eluate is very low, less than 0.05% or one hit per 2000 compounds, the average number of hits per eluate is less than one hit per large mixture. If the ESI-MS assay is performed using HPLC, sample volume due to pooling many eluates is not a problem since the sample is concentrated on the column. The main concern will be the amount of protein and its effect on the mass spectra. Often this is not a problem due to the low levels of protein used in the assays. However, the protein can be easily removed as discussed below in Section 2.2.10.
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