Retentate chromatography and protein chip arrays with applications in biology and medicine
First Claim
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1. A method for determining whether an agent modulates binding between a receptor/ligand pair that specifically bind to each other, the method comprising the steps of:
- a) providing a mass spectrometry probe, the probe comprising a substrate having a surface and a receptor or ligand of said receptor/ligand pair docked to a surface of the substrate through an adsorbent;
b) exposing the receptor or the ligand docked to the substrate to its binding partner and to said agent under an elution conditions that allows for binding between the receptor and the ligand;
c) removing unbound binding partner from the surface of the substrate;
d) measuring the amount of binding partner retained on the docked receptor or ligand in the presence and absence of the agent by laser desorption mass spectrometry of any retained binding partner from the surface of the substrate; and
e) determining whether the agent modulates binding by comparing the measured amount of binding between the receptor and the ligand in the presence and absence of the agent whereby a difference between the measured amount of binding between the receptor and ligand in the presence and absence of the agent indicates that the agent modulates binding between the receptor/ligand pair.
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Abstract
This invention provides methods of retentate chromatography for resolving analytes in a sample. The methods involve adsorbing the analytes to a substrate under a plurality of different selectivity conditions, and detecting the analytes retained on the substrate by desorption spectrometry. The methods are useful in biology and medicine, including clinical diagnostics and drug discovery.
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Citations
13 Claims
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1. A method for determining whether an agent modulates binding between a receptor/ligand pair that specifically bind to each other, the method comprising the steps of:
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a) providing a mass spectrometry probe, the probe comprising a substrate having a surface and a receptor or ligand of said receptor/ligand pair docked to a surface of the substrate through an adsorbent;
b) exposing the receptor or the ligand docked to the substrate to its binding partner and to said agent under an elution conditions that allows for binding between the receptor and the ligand;
c) removing unbound binding partner from the surface of the substrate;
d) measuring the amount of binding partner retained on the docked receptor or ligand in the presence and absence of the agent by laser desorption mass spectrometry of any retained binding partner from the surface of the substrate; and
e) determining whether the agent modulates binding by comparing the measured amount of binding between the receptor and the ligand in the presence and absence of the agent whereby a difference between the measured amount of binding between the receptor and ligand in the presence and absence of the agent indicates that the agent modulates binding between the receptor/ligand pair. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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Specification