US 8,525,104 B2Granted

Methods for direct biomolecule identification by matrix-assisted laser desorption ionization (MALDI) mass spectrometry

AssigneeNew York University

InventorsPaul Pevsner, Frederick Naftolin, Douglas C. Miller, Dean Hillman, Brian K. Stall, Steven M. Wishnies

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Abstract

The present invention relates to the use of post source decay (PSD) or collision induced dissociation (CID) direct tissue (DT) MALDI-TOF or DT-MALDI-TOF-TOF mass spectrographic identification of biological molecules in a tissue or cellular sample without the need for further protein extraction. This method provides for studying cells or tissues by direct tissue MALDI (DT-MALDI), thereby substituting in situ protein release for further protein extraction. Mass/intensity data was processed with Mascot© software interrogation of the NCBI database. These results are proof of principle that DT-MALDI, combined with bioinformatics, can directly identify proteins in cells and tissues from their mass spectra.

Claims17

Dependent claims

A method for analyzing the biological molecule content of a tissue sample obtained from normal tissue or abnormal/diseased tissue in situ, wherein the method provides for a level of detection of the biological molecules in the sample in an amount ranging from about 1 attamol to about 10 attamols, comprising:

a) collecting a sample of tissue from a subject into a first solution effective to maintain integrity of the biological molecule;
b) treating the sample with a second solution comprising one or more enzymes, or chemicals, effective to dissociate the tissue sample or of digesting the dissociated tissue sample into smaller fragments;
c) treating the preparation from step b) with a matrix assisted laser desorption ionization imaging (MALDI) matrix solution;
d) analyzing the preparation from step c) by direct tissue (DT)-matrix assisted laser desorption ionization imaging (MALDI)-time of flight (TOF) measurement or DT-MALDI-TOF-TOF measurement;
e) creating a data file utilizing the information from step d);
f) entering the data from the data file of step e) into an external database to create a signature map for the tissue from which the data was obtained;
and
g) comparing the results from step d) with a signature map for normal tissue, wherein said normal tissue corresponds to, or is of the same tissue type, as the tissue from which the sample was obtained.

5 more elements

Dependencies are inferred from the claim text.

Facts

Application: 12/799,576Filed 27 Apr 2010 · continuation
Priority: 30 Sep 2005
Published: US 2011/0062320 A117 Mar 2011
Granted: 3 Sep 2013
Term ends: Not estimatedThe source records don’t support an estimate.

Classification5 codes

G01N 33/6851
Methods of protein analysis involving mass spectrometry: methods of protein analysis involving laser desorption ionisation mass spectrometry
G01N 2800/52
Detection or diagnosis of diseases: predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in…
H01J 49/164
Using photoionisation, e.g. by laser: laser desorption/ionisation, e.g. matrix-assisted laser desorption/ionisation [MALDI]
G01N
Investigating or analysing materials by determining their chemical or physical properties

Family

Application60/722,206Us provisional application · filed 30 Sep 2005
Application60/784,016Us provisional application · filed 20 Mar 2006
Application11/541,380Parent of this continuation · filed 29 Sep 2006
US 2011/0062320 A1Pre-grant publication · published 17 Mar 2011
US 7,714,276 B2Parent of this continuation
US 2007/0114375 A1Published application
US 8,525,104 B2This patent

From the US filing history. Foreign family members aren’t listed.