Retentate chromatography and protein chip arrays with applications in biology and medicine
First Claim
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1. A method for high information resolution of at least one analyte in a sample comprising the steps of:
- a) exposing the analyte to at least two different selectivity conditions, each selectivity condition defined by the combination of an adsorbent and an eluant, to allow retention of the analyte by the adsorbent; and
b) detecting retained analyte under the different selectivity conditions by desorption spectrometry;
whereby detection of retained analyte under the different selectivity conditions provides a high information resolution of the analyte.
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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
31 Claims
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1. A method for high information resolution of at least one analyte in a sample comprising the steps of:
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a) exposing the analyte to at least two different selectivity conditions, each selectivity condition defined by the combination of an adsorbent and an eluant, to allow retention of the analyte by the adsorbent; and
b) detecting retained analyte under the different selectivity conditions by desorption spectrometry;
whereby detection of retained analyte under the different selectivity conditions provides a high information resolution of the analyte. - 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)
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24. A substrate for desorption spectrometry comprising an adsorbent whose binding characteristics vary in a gradient along one or more linear axes.
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25. A method for preparative purification of an analyte from an impure sample comprising the steps of:
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a) exposing the sample to a substrate under a plurality of different selectivity conditions;
detecting retained analyte under the different selectivity conditions by desorption spectrometry; and
identifying selectivity conditions under which the analyte is retained;
b) purifying the analyte by repeating, for a plurality of different identified selectivity conditions, a sequence of steps comprising;
i) exposing the sample to an adsorbent under the identified selectivity condition to allow retention of the analyte by the adsorbent;
ii) separating the analyte from an impurity that is not retained by the substrate; and
iii) collecting the analyte from the adsorbent;
whereby the analyte is purified.
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26. A method for selecting identity candidates for an analyte protein comprising the steps of:
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a) determining a value set specifying match parameters for at least a first and second physico-chemical characteristic of a protein analyte in a sample by i) exposing the analyte to a plurality of different selectivity conditions, wherein adsorption of the protein analyte to the substrate is mediated by a basis of attraction that identifies a physico-chemical characteristic of the protein analyte; and
ii) detecting retained analyte under the different selectivity conditions by desorption spectrometry; and
b) performing, in a programmable digital computer, the steps of;
i) accessing a database comprising, for each member of a set of reference polypeptides, a value set specifying at least a first and second physico-chemical characteristic of the reference polypeptides;
ii) accessing the value set specifying the physico-chemical characteristics of the protein analyte;
iii) sorting from the database, reference polypeptides having value sets within the match parameters;
whereby the sorted reference polypeptides provide identity candidates for the protein analyte and unsorted reference polypeptides provide non-identity candidates for the protein analyte. - View Dependent Claims (27, 28, 29)
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30. A method of detecting an enzyme in a sample comprising the steps of:
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a) providing a solid phase comprising an adsorbent and an enzyme substrate bound to the adsorbent, wherein the activity of the enzyme on the enzyme substrate produces a product having a characteristic molecular mass;
b) exposing the substrate to the sample; and
c) detecting the product by desorption spectrometry;
whereby detecting the product provides a detection of the enzyme. - View Dependent Claims (31)
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Specification