Tagged Ligands For Enrichment of Rare Analytes From A Mixed Sample
First Claim
1. A method for enriching rare cells from a cellular sample comprising:
- contacting in solution said cellular sample with affinity-tagged ligands (ATLs) each comprising;
a first ligand linked to an affinity tag, wherein said ligand selectively binds a cellular marker of said rare cells and said affinity tag can be selectively captured by a capture moiety, wherein said affinity tags do not comprise a magnetic particle; and
flowing said sample through a microfluidic device comprising said capture moiety to selectively retain ATL-bound cells.
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Abstract
Method of enriching specific cells from cellular samples are disclosed, comprising contacting in solution a cellular sample with affinity-tagged ligands (ATLs) each comprising a first ligand linked to an affinity tag, wherein the ligand selectively binds a cellular marker of the rare cells and the affinity tag can be selectively captured by a capture moiety, wherein the affinity tags do not comprise a magnetic particle; and flowing the sample through a microfluidic device comprising the capture moiety to selectively retain ATL-bound cells. Methods for enriching circulating tumor cells, and devices for enriching specific cells from cellular samples are also disclosed.
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Citations
28 Claims
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1. A method for enriching rare cells from a cellular sample comprising:
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contacting in solution said cellular sample with affinity-tagged ligands (ATLs) each comprising; a first ligand linked to an affinity tag, wherein said ligand selectively binds a cellular marker of said rare cells and said affinity tag can be selectively captured by a capture moiety, wherein said affinity tags do not comprise a magnetic particle; and flowing said sample through a microfluidic device comprising said capture moiety to selectively retain ATL-bound cells. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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2. A method for enriching rare cells from a cellular sample comprising:
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contacting in solution said cellular sample with affinity-tagged ligands (ATLs) each comprising; a ligand linked to a plurality of affinity tags, wherein said ligand selectively binds a cellular marker of said rare cells and said affinity tag can be selectively captured by a capture moiety and wherein the ratio of ligand;
affinity tag of said ATL'"'"'s is less than 1;
1flowing said sample through a microfluidic device comprising said capture moiety to selectively retain ATL-bound cells.
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11. A method for enriching rare analytes from a sample comprising:
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contacting in solution said sample with a plurality of affinity-tagged ligands (ATLs) wherein said mixture of ATL'"'"'s comprises; a first ATL comprising a first ligand that selectively binds a first marker of said rare analytes, wherein said first ligand is linked to a first affinity tag that is selectively captured by a first capture moiety; and a second ATL comprising a second ligand that selectively binds a second marker of said rare analytes, wherein said second ligand is linked to a second affinity tag, wherein said second affinity tag is selectively captured by said first capture moiety; and contacting said sample with said capture moiety to selectively enrich said rare analytes. - View Dependent Claims (12, 13, 14)
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- 25. A reagent comprising a plurality of affinity tagged ligands each of which comprises a ligand that selectively binds a different cancer cellular marker linked to an identical affinity tag that is selectively captured by a capture moiety.
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28. A microfluidic device comprising an array of obstacles, wherein the obstacles are functionalized with one or more capture moieties selected from the group comprising EGF, EGF-peptide, EPI-peptide, folate or DBA.
Specification