Deep MALDI TOF mass spectrometry of complex biological samples, e.g., serum, and uses thereof
A method of analyzing a biological sample, for example serum or other blood-based samples, using a MALDI-TOF mass spectrometer instrument is described. The method includes the steps of applying the sample to a sample spot on a MALDI-TOF sample plate and directing more than 20,000 laser shots to the sample at the sample spot and collecting mass-spectral data from the instrument. In some embodiments at least 100,000 laser shots and even 500,000 shots are directed onto the sample. It has been discovered that this approach, referred to as “deep-MALDI”, leads to a reduction in the noise level in the mass spectra and that a significant amount of additional spectral information can be obtained from the sample. Moreover, peaks visible at lower number of shots become better defined and allow for more reliable comparisons between samples.
1. A method of performing MALDI-TOF mass spectrometry on a sample, comprising the steps of:
A) applying the sample to multiple predefined sample spots of a MALDI plate;
B) raster scanning the multiple predefined sample spots of the MALDI plate with a laser in accordance with a raster file defining a pattern for raster scanning of a spot of a MALDI sample plate at a multitude of noncontiguous X-Y locations and acquiring mass spectra of the sample from each of the X-Y locations of the spots, wherein the raster scanning comprises directing a multitude of laser shots to each of the noncontiguous X-Y locations;
C) after performing normalization and alignment operations, summing the mass spectra acquired in step B);
wherein the sample is subject to a total of at least 100,000 laser shots in the raster scanning of the multiple predefined sample spots.
2. The method of claim 1 , wherein the sample is in the form of an unfractionated blood-based sample from a human.
3. The method of claim 1 , wherein the sample is in the form of a fractionated blood-based sample from a human.
4. The method of claim 1 , further comprising the step of accepting or rejecting transient spectra acquired during step B) by applying a filtering criteria to the transient spectrum.
5. The method of claim 4 , wherein the filtering criteria comprises determining whether the transient spectra have a peak in a predefined m/z range and if not rejecting the transient spectra.
6. The method of claim 1 , wherein at least 100,000 mass spectra of the sample are acquired from each of the multiple spots.