Retentate chromatography and protein chip arrays with applications in biology
First Claim
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1. A method for preparing a substrate for detecting at least one analyte in a sample comprising the steps of:
- a) exposing the sample to at least two different selectivity conditions, each selectivity condition defmed by the combination of an adsorbent and an eluant, to allow retention of the analyte by the adsorbent;
b) identifying by desorption spectrometry at least one selectivity condition under which the analyte is retained; and
c) preparing a substrate comprising at least one adsorbent of an identified selectivity condition.
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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
30 Claims
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1. A method for preparing a substrate for detecting at least one analyte in a sample comprising the steps of:
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a) exposing the sample to at least two different selectivity conditions, each selectivity condition defmed by the combination of an adsorbent and an eluant, to allow retention of the analyte by the adsorbent;
b) identifying by desorption spectrometry at least one selectivity condition under which the analyte is retained; and
c) preparing a substrate comprising at least one adsorbent of an identified selectivity condition. - View Dependent Claims (2, 3)
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4. A method for progressively identifying a selectivity condition with improved resolution for an analyte in a sample comprising the steps of:
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(a) identify a selectivity condition that retains an analyte in a sample by;
(i) exposing a sample to a set of selectivity conditions, each selectivity condition defined by at least one binding characteristic and at least one elution characteristic;
(ii) detecting analyte retained under each selectivity condition by desorption spectrometry; and
(iii) identifying a selectivity condition that retains the analyte; and
(b) identifying a selectivity condition with improved resolution for the analyte by;
(i) selecting at least one binding characteristic or elution characteristic from the identified selectivity condition and adding it to a selectivity characteristic constant set;
(ii) exposing the sample to a modified set of selectivity conditions wherein each selectivity condition in the modified set comprises (1) the selectivity characteristics in the constant set and (2) a binding characteristic or elution characteristic that is not in the constant set; and
(iii) identifying a selectivity condition from the modified set by desorption spectrometry that retains the analyte with improved resolution compared with a prior identified selectivity condition. - View Dependent Claims (5, 6, 7, 8)
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9. A method for determining whether an analyte is differentially present in a first and second biological sample comprising the steps of:
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a) determining a first retention map for the analyte in the first sample for at least one selectivity condition;
b) determining a second retention map for the analyte in the second sample for the same selectivity condition; and
c) detecting a difference between the first and the second retention maps;
whereby a difference in the retention maps provides a determination that the analyte is differentially present in first and second samples. - View Dependent Claims (10, 11, 12, 13, 14, 15, 16, 17, 18, 20)
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19. A method of diagnosing in a subject a disease characterized by at least one diagnostic marker comprising the steps of:
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a) providing a substrate for use in desorption spectrometry that comprises at least one addressable location, each addressable location comprising an adsorbent that resolves at least one of the diagnostic markers under an elution condition;
b) exposing the substrate to a biological sample from the subject under the elution condition to allow retention of the diagnostic marker; and
c) detecting retained diagnostic marker by desorption spectrometry;
whereby detecting retained diagnostic marker provides a diagnosis of the disease.
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- 21. A kit for detecting an analyte in a sample comprising (1) a substrate for use in desorption spectrometry that comprises at least one addressable location, each addressable location comprising an adsorbent that resolves an analyte under a selectivity condition comprising the adsorbent and an eluant, and (2) the eluant or instructions for exposing the sample to the selectivity condition.
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28. A substrate for desorption spectrometry comprising at least one adsorbent in at least one addressable location wherein the at least one adsorbent resolves a plurality of diagnostic markers for a pathological condition from a patient sample.
Specification