Tagged ligands for enrichment of rare analytes from a mixed sample
AssigneeGPB Scientific, LLC
InventorsAllyn Forsyth, Helen Barnes
Life of this patent
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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.
Claims20
A method for enriching rare analytes from a sample, comprising:
- a) contacting said sample with an affinity-tagged ligand (ATL) complex wherein said ATL complex comprises:
- i) an affinity tag comprising a magnetic particle;
- and
- ii) at least a first ligand and a second ligand different from said first ligand, wherein said first ligand and said second ligand are linked to said affinity tag;
- and wherein said first ligand and said second ligand selectively bind different markers of said rare analytes, thereby labeling said rare analytes with said ATL complex;
- b) flowing said sample comprising said ATL complex-labeled rare analytes through a microfluidic enrichment device comprising an array of obstacles, wherein said obstacles are arranged in rows offset from other obstacles to create restricted and expanded gaps and to thereby laterally displace ATL complex-labeled rare analytes based on their hydrodynamic size, and wherein at least some of said obstacles are coated or treated with one or more capture moieties that capture the labeled rare analytes;
- wherein said capture moieties comprise a magnet or magnetized substrate that interacts with the magnetic particle of the affinity tag, to selectively enrich said labeled rare analytes.
A method for enriching leukocytes or stem cells in a sample comprising:
- a. contacting said sample with an affinity-tagged ligand (ATL) complex, wherein said ATL complex comprises:
- i. an affinity tag comprising a magnetic particle;
- and
- ii. at least a first ligand and a second ligand different from said first ligand, wherein said first ligand and said second ligand are linked to said affinity tag, and wherein said first ligand and said second ligand selectively bind different markers of said leukocytes or stem cells, thereby labeling said leukocytes or stem cells with said ATL complex;
- b. flowing said sample comprising said ATL complex-labeled leukocytes or stem cells through a microfluidic device comprising an array of obstacles, wherein said obstacles are arranged in rows offset from other obstacles and that laterally displace the ATL complex-labeled leukocytes or stem cells based on their hydrodynamic size;
- and
- c. flowing said sample comprising said ATL complex-labeled specific cells that have been flowed through the microfluidic device in step b) through an enrichment device comprising a surface functionalized with a capture moiety, wherein said capture moiety comprises a magnet or magnetized substrate that interacts with the magnetic particle of the affinity tag on the ATL complex-labeled leukocytes or stem cells to selectively enrich said ATL complex-labeled specific cells.
Dependencies are inferred from the claim text.