Retentate chromatography and protein chip arrays with applications in biology and medicine
First Claim
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1. A method for detecting polypeptide analytes in a sample comprising the steps of:
- a) exposing the sample to a plurality of different selectivity conditions, each selectivity condition defined by a combination of an adsorbent and an eluant, i) wherein said plurality of different selectivity conditions comprises at least two different adsorbents having different bases of attraction, wherein the bases of attraction are selected from the group consisting of a hydrophobic interaction, a hydrophilic interaction, an anionic interaction, a cationic interaction and a coordinate covalent interaction; and
ii) wherein exposing comprises (I) contacting the sample with an adsorbent bound to a substrate; and
(II) washing unbound polypeptides from the adsorbent with the eluant to allow retention of polypeptide analytes on the adsorbent; and
b) detecting retained polypeptide analytes on each adsorbent by laser desorption mass spectrometry.
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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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27 Claims
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1. A method for detecting polypeptide analytes in a sample comprising the steps of:
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a) exposing the sample to a plurality of different selectivity conditions, each selectivity condition defined by a combination of an adsorbent and an eluant, i) wherein said plurality of different selectivity conditions comprises at least two different adsorbents having different bases of attraction, wherein the bases of attraction are selected from the group consisting of a hydrophobic interaction, a hydrophilic interaction, an anionic interaction, a cationic interaction and a coordinate covalent interaction; and
ii) wherein exposing comprises (I) contacting the sample with an adsorbent bound to a substrate; and
(II) washing unbound polypeptides from the adsorbent with the eluant to allow retention of polypeptide analytes on the adsorbent; and
b) detecting retained polypeptide analytes on each adsorbent by laser desorption mass spectrometry. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
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Specification