Retentate chromatography and protein chip arrays with applications in biology and medicine
First Claim
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1. A method for preparative purification of an analyte from an impure sample comprising the steps of:
- a) identifying a selectivity condition under which the analyte is retained by;
i) exposing the sample to a substrate under a plurality of different selectivity conditions, each selectivity condition defined by a combination of an adsorbent and an eluant, wherein exposing comprises (I) contacting the sample with an adsorbent bound to a substrate; and
(II) washing unbound analyte from the adsorbent with the eluant to allow retention of analytes on the adsorbent; and
ii) detecting a selectivity condition under which the analyte is retained by laser desorption mass spectrometry; and
b) purifying the analyte by;
i) contacting the sample with a chromatography column comprising an adsorbent identified using the process recited in step (a) to retain the analyte under the identified selectivity condition;
ii) washing unbound material from the chromatography column; and
iii) eluting the analyte from the chromatography column;
whereby the analyte is purified.
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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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10 Claims
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1. A method for preparative purification of an analyte from an impure sample comprising the steps of:
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a) identifying a selectivity condition under which the analyte is retained by;
i) exposing the sample to a substrate under a plurality of different selectivity conditions, each selectivity condition defined by a combination of an adsorbent and an eluant, wherein exposing comprises (I) contacting the sample with an adsorbent bound to a substrate; and
(II) washing unbound analyte from the adsorbent with the eluant to allow retention of analytes on the adsorbent; and
ii) detecting a selectivity condition under which the analyte is retained by laser desorption mass spectrometry; and
b) purifying the analyte by;
i) contacting the sample with a chromatography column comprising an adsorbent identified using the process recited in step (a) to retain the analyte under the identified selectivity condition;
ii) washing unbound material from the chromatography column; and
iii) eluting the analyte from the chromatography column;
whereby the analyte is purified. - View Dependent Claims (2, 3, 4, 5)
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6. A method of preparative purification of an analyte comprising:
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(a) determining at least one selectivity condition that retains an analyte, wherein a selectivity condition is defined by a combination of an adsorbent and an eluant, by;
(i) contacting a sample comprising the analyte with an adsorbent bound to a substrate, (ii) washing unbound material from the adsorbent with the eluant; and
(iii) determining whether the adsorbent retained the analyte by laser desorption mass spectrometry from the substrate; and
(b) purifying the analyte from a sample by;
(i) contacting a sample comprising the analyte in a chromatography column with an adsorbent identified using the process recited in step (a) to retain the analyte, (ii) washing unbound material from the column with an eluant, whereby the analyte is retained in the column; and
(iii) eluting the analyte from the column;
whereby the analyte is purified. - View Dependent Claims (7, 8, 9, 10)
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Specification