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Deep-maldi tof mass spectrometry of complex biological samples, e.g., serum, and uses thereof

A technology for biological samples and samples, which is used in mass spectrometers, dynamic spectrometers, particle imaging spectroscopy, etc.

Inactive Publication Date: 2015-02-25
BIODESIX
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, it is generally believed that MALDI-TOF MS of serum can only be used for high-abundance proteins in the micromole per liter range

Method used

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  • Deep-maldi tof mass spectrometry of complex biological samples, e.g., serum, and uses thereof
  • Deep-maldi tof mass spectrometry of complex biological samples, e.g., serum, and uses thereof
  • Deep-maldi tof mass spectrometry of complex biological samples, e.g., serum, and uses thereof

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Embodiment

[0073] In one embodiment of this method, spectra can be collected from 5 million shots from multiple spots of the same serum on a MALDI plate using techniques previously described using manually or automatically generated rasters for scanning the multiple spots . In this method, uniform spots of individual samples on MALDI plates are preferably obtained reproducibly. This can be achieved using the method described in the present invention.

[0074] 1. Spot the diluted serum onto the MALDI target plate.

[0075] Process:

[0076] Serum was diluted 1:10 with HPLC grade water and vortexed. Samples were mixed 1:1 (v / v) with matrix (20 mg / ml sinapinic acid in 50% ACN / 0.1% TFA) in a 0.5 ml centrifuge tube and vortexed. Spot 4 [mu]l of matrix / sample mixture onto one or more spots on the MALDI target.

[0077] Thirty-six points (positions) in the MALDI plate were used in this example:

[0078] Tube 1: Spotted at positions E13, E14 and E15 of the MALDI plate (see Figure 2A )

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Abstract

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.

Description

[0001] Cross References to Related Applications [0002] This application claims priority under 35 U.S.C. §119 to U.S. Provisional Application Serial No. 61 / 652,394, filed May 29, 2012, the contents of which are incorporated herein by reference. [0003] background [0004] The present disclosure relates to the fields of mass spectrometry, biomarker discovery, assay development, and clinical testing. Background technique [0005] In MALDI (matrix assisted laser desorption ionization, matrix assisted laser desorption ionization) TOF (time-of-flight, time-of-flight) mass spectrometry, the sample / matrix mixture is placed on a metal plate (called a MALDI plate) at defined positions ("point" or "sample point" in this article). A laser beam is directed to a location on the spot for a very short instant (called a "shot"), causing desorption and ionization of molecules or other components of the sample. This sample component "flies" to the particle detector. The instrument measur...

Claims

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Application Information

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IPC IPC(8): H01J49/16H01J49/00G01N33/487
CPCG01N33/6851H01J49/164G01N33/487H01J49/0004H01J49/40
Inventor 海因里希·罗德S·阿斯梅拉希J·艾伦马克西姆·齐平
Owner BIODESIX
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