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US 2014/0235471 A1Published application
Global Proteomic Screening Of Random Bead Arrays Using Mass Spectrometry Imaging
AssigneeAmberGen, Inc.
InventorsVladislav B. Bergo, Mark J. Lim, Kenneth J. Rothschild
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Abstract
Methods for proteomic screening on random protein-bead arrays by mass spec is described. Photocleavable mass tags are utilized to code a protein library (bait molecules) displayed on beads randomly arrayed in an array substrate. A library of probes (prey) can be mixed with the protein-bead array to query the array. Because mass spec can detect multiple mass tags, it is possible to rapidly identify all of the interactions resulting from this mixing.
Claims26
A method of detecting the interaction of prey molecules with bait molecules on an array of beads, comprising:
- a. providing i) a mixture comprising first and second beads, said first beads comprising first bait molecules, said second beads comprising second bait molecules, wherein said first and second bait molecules are different;
- and ii) prey molecules, wherein said prey molecules comprise a detectable label;
- b. contacting said first and second beads with a solution comprising a population of said prey molecules under conditions wherein said prey molecules can bind to a bait molecule;
- c. making an array with said beads;
- d. imaging said beads in said array for the presence of said detectable label to detect interaction of prey molecules with bait molecules;
- and
- e. subjecting one or more beads detected in said array in step d) to MALDI-MS measurements.
4 more elements
A method of detecting the interaction of prey molecules with bait molecules on an array of beads, comprising:
- a. providing i) a mixture comprising first and second beads, said first beads comprising first bait molecules, said second beads comprising second bait molecules, wherein said first and second bait molecules are different;
- ii) prey molecules;
- and iii) detectable labels capable of being coupled to said prey molecules;
- b. contacting said first and second beads with a first solution comprising a population of said prey molecules;
- c. contacting said first and second beads with a second solution comprising said detectable labels;
- d. making an array with said beads;
- e. imaging said beads in said array for the presence of said detectable labels;
- and
- f. subjecting one or more beads in said array identified in step e to MALDI-MS measurements.
6 more elements
A method of making and decoding an array, comprising:
- a) providing i) a mixture comprising first and second beads, said first beads comprising first bait molecules, said second beads comprising second bait molecules, wherein said first and second bait molecules are different;
- and ii) a surface;
- b) making an array with said beads on said surface;
- c) imaging said array;
- and
- d) subjecting said array to MALDI-MS analysis under conditions wherein the identity of said bait on at least a portion of said beads is determined.
3 more elements
A method of detecting the interaction of prey molecules with bait molecules on an array of beads, comprising:
- a. providing i) a mixture comprising first and second beads, said first beads comprising first bait molecules, said second beads comprising second bait molecules, wherein said first and second bait molecules are different;
- and ii) prey molecules, wherein said prey molecules comprise a detectable label;
- b. contacting said first and second beads with a solution comprising a population of said prey molecules under conditions wherein said prey molecules can bind to a bait molecule;
- c. making an array with said beads;
- d. imaging said beads in said array for the presence of said detectable label to detect positive beads indicating an interaction of prey molecules with bait molecules;
- e. subjecting at least a portion of said array to MALDI-MS imaging;
- and
- f. synchronizing images detected in steps d) and e) to decode beads that have bait-prey interactions, wherein the position of the positive beads detected in step d) is used to direct the MALDI-MS spectrometer to the position of these positive beads.
5 more elements
A method of detecting the interaction of prey molecules with bait molecules on an array of beads, comprising:
- a. providing i) a mixture comprising first and second beads, said first beads comprising first bait molecules, said second beads comprising second bait molecules, wherein said first and second bait molecules are different;
- ii) prey molecules;
- and iii) detectable labels capable of being coupled to said prey molecules;
- b. contacting said first and second beads with a solution comprising a population of said prey molecules;
- c. contacting said first and second beads with a solution comprising said detectable labels;
- d. making an array with said beads;
- e. imaging said beads in said array for the presence of said detectable label to detect positive beads in a position indicating an interaction of prey molecules with bait molecules;
- f. subjecting at least a portion of said array to MALDI-MS imaging;
- and
- g. synchronizing images detected in steps e) and f) to decode beads that have bait-prey interactions, wherein the position of the positive beads detected in step e) is used to direct the MALDI-MS spectrometer to the position of these positive beads.
7 more elements
A method of detecting the interaction of prey molecules with bait molecules on an array of beads, comprising:
- a. providing i) a mixture comprising first and second beads, said first beads comprising first bait molecules, said second beads comprising second bait molecules, wherein said first and second bait molecules are different;
- and ii) prey molecules, wherein said prey molecules comprise a detectable label;
- b. contacting said first and second beads with a solution comprising a population of said prey molecules under conditions wherein said prey molecules can bind to a bait molecule;
- c. making an array with said beads;
- d. imaging said beads in said array for the presence of said detectable label to detect the position of positive beads indicating an interaction of prey molecules with bait molecules;
- and
- e. using the position of the beads detected in step d) to direct the MALDI-MS spectrometer to the position of these positive beads.
4 more elements
A method of detecting the interaction of prey molecules with bait molecules on an array of beads, comprising:
- a. providing i) a mixture comprising first and second beads, said first beads comprising first bait molecules, said second beads comprising second bait molecules, wherein said first and second bait molecules are different;
- ii) prey molecules;
- and iii) detectable labels capable of being coupled to said prey molecules;
- b. contacting said first and second beads with a solution comprising a population of said prey molecules;
- c. contacting said first and second beads with a solution comprising said detectable labels;
- d. making an array with said beads;
- e. imaging said beads in said array for the presence of said detectable label to detect the position of positive beads indicating an interaction of prey molecules with bait molecules;
- f. using the position of the beads detected in step e) to direct the MALDI-MS spectrometer to the position of these positive beads.
5 more elements
Dependencies are inferred from the claim text.