Glycan Analysis of Proteins in Cells
AssigneesMUSC Foundation for Research Development, Van Andel Research Institute
InventorsAnand Mehta, Richard R. Drake, Brian Haab, Peggi M. Angel
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Abstract
The present invention provides methods and compositions for glycan analysis of complex solutions, including proteins and cells in a biological sample. The method includes the preparation of substrates for the capture of proteins and cells for multiplexed analysis. Cells and proteins may be captured by antibody arrays, culture, or direct deposition. The invention further relates to the use of protein and cell glycan analysis in the diagnosis and screening of disease states and disease progression.
Claims31
A method for glycan analysis of at least one sample, the method comprising the steps of:
- providing a substrate having a surface spotted with a plurality of different antibodies positioned at discrete locations on the substrate, wherein the plurality of different antibodies bind to a plurality of different proteins;
- incubating the substrate in a blocking solution;
- incubating the substrate in at least one sample, wherein the at least one sample comprises at least one protein solution derived from a biological sample, wherein the proteins in the sample bind to the plurality of different antibodies, thereby capturing the proteins;
- spraying the substrate with an enzymatic releasing solution to release glycans from the captured proteins;
- and
- scanning the substrate by mass spectrometry to detect and identify the glycans present at each discrete location, thereby identifying a plurality of glycoproteins in the sample.
(canceled)
A method for glycan analysis of at least one population of cells, the method comprising the steps of:
- adhering at least one population of cells to a surface of a substrate;
- fixing and rinsing the at least one population of cells;
- spraying the substrate with an enzymatic releasing solution to release glycans from the captured cells;
- and
- scanning the substrate by mass spectrometry to detect and identify the presence of glycans;
- wherein the at least one population of cells comprises cultured or captured cells.
A kit for glycan analysis of protein samples, comprising:
- at least one substrate, each substrate having a surface spotted with a plurality of antibodies;
- at least one blocking solution;
- at least one enzymatic releasing solution;
- and
- at least one MALDI matrix material.
Dependencies are inferred from the claim text.