High-Throughput Axial MALDI-TOF MS Using a 2 kHz Repetition Rate Laser
| Authors | |
|---|---|
| Year of publication | 2006 |
| Type | Article in Periodical |
| Magazine / Source | Analytical Chemistry |
| MU Faculty or unit | |
| Citation | |
| Field | Analytic chemistry |
| Keywords | MALDI; TOF; mass spectrometry; time of flight; high-throughput |
| Description | A high-throughput axial MALDI-TOF mass spectrometer utilizing a laser with a 2 kHz pulse repetition-rate was constructed and tested. This fast mass spectrometer provided a data acquisition rate 10 times faster than a commercially available (200 Hz) axial mass spectrometer, while maintaining comparable limits of detection (200 amole of Glu fib peptide). Mass resolution, only slightly less than the commercial instrument (10,000 vs. 14,000), was sufficient for baseline resolution of isotopic clusters of peptides with m/z <2700. A new method of mass calibration, which combined a limited number of internal and external standards, provided the same 15 ppm mass accuracy over the entire sample plate on either instrument. Implementing the 2 kHz laser required a faster data acquisition system and high-voltage pulse electronics, together with a novel strategy for rapid sample plate movements during acquisition, to achieve a sample analysis rate of up to 2 spots per second (with 800 shots/spot). The overall performance of the fast MALDI-TOF MS instrument was demonstrated by the acquisition, in 12 minutes, of an LC-MS dataset from a plate of 650 fractions collected during LC separation of an 16O/18O differentially-labeled proteomic sample of a tryptic digest of an E.coli lysate. |
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