Process for detection of fetal cells
AssigneeMenarini Silicon Biosystems S.p.A.
InventorsAnna Doffini, Chiara Maranta, Chiara Mangano, Emilia Dora Giovannone, Thomas J. Musci
Life of this patent
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Abstract
Provided herein is a method of isolating fetal cells from a sample from a pregnant subject. The method utilizes magnetic particles conjugated to anti-EGFR or a combination of anti-EGFR and anti-CD105 antibodies to create an enriched sample and then labeled antibodies for fetal cell markers to isolate different fetal cell types in the sample from the pregnant subject. Said isolated fetal cells can then be analyzed via gene sequencing or genomic or genetic analysis.
Claims44
A method for isolating fetal cells from a sample from a pregnant subject, comprising:
- (a) contacting the sample comprising a plurality of cells with a plurality of magnetic particles, the magnetic particles conjugated to an anti-epidermal growth factor receptor (EGFR) antibody;
- (b) isolating cells bound to the anti-EGFR antibody by subjecting the sample to a magnetic field, thereby producing an enriched sample;
- (c) contacting the enriched sample with a first labeled antibody that binds to a cytokeratin (CK);
- and
- (d) isolating a fetal cell that is bound to the first labeled antibody;
- wherein the fetal cell is a fetal nucleated red blood cell (fnRBC) or a trophoblast.
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A method for isolating fetal cells from a sample from a pregnant subject, comprising:
- (a) contacting the sample comprising a plurality of cells with a plurality of magnetic particles, wherein each magnetic particle in the plurality is conjugated to an anti-EGFR antibody;
- (b) subjecting the sample to a magnetic field;
- (c) isolating cells bound to the magnetic particles to produce an enriched sample;
- (d) contacting the enriched sample with:
- (i) an antibody that binds to a cytokeratin (CK) and is conjugated to a fluorescent label,
- (ii) an antibody that binds to CD45 and is conjugated to a fluorescent label, and
- (iii) a nuclear stain;
- and
- (d) isolating a fetal cell using fluorescence activated cell sorting (FACS) or DEPArray, wherein the fetal cell is positive for the CK and nuclear stain and negative for CD45;
- wherein the fetal cell is a fetal nucleated red blood cell (fnRBC) or a trophoblast.
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A method for isolating fetal cells from a sample from a pregnant subject, comprising:
- (a) contacting the sample comprising a plurality of cells with a plurality of magnetic particles, wherein the plurality of magnetic particles comprises:
- (i) magnetic particles that are conjugated to an anti-epidermal growth factor receptor (EGFR) antibody and an anti-CD105 antibody, or
- (ii) a combination of magnetic particles that are conjugated to an anti-EGFR antibody and magnetic particles that are conjugated to an anti-CD105 antibody;
- (b) isolating cells bound to the first antibody or the second antibody by subjecting the sample to a magnetic field, thereby producing an enriched sample;
- (c) contacting the enriched sample with a first labeled antibody that binds to a cytokeratin (CK);
- and
- (d) isolating a fetal cell that is bound to the first labeled antibody;
- wherein the fetal cell is a fetal nucleated red blood cell (fnRBC) or a trophoblast.
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A method for isolating fetal cells from a sample from a pregnant subject, comprising:
- (a) contacting the sample comprising a plurality of cells with a plurality of magnetic particles, wherein the plurality of magnetic particles comprises:
- (i) magnetic particles that are conjugated to an anti-epidermal growth factor receptor (EGFR) antibody and an anti-CD105 antibody, or
- (ii) a combination of magnetic particles that are conjugated to an anti-EGFR antibody and magnetic particles that are conjugated to an anti-CD105 antibody;
- (b) subjecting the sample to a magnetic field;
- (c) isolating cells bound to the magnetic particles to produce an enriched sample;
- (d) contacting the enriched sample with:
- (i) an antibody that binds to a cytokeratin (CK) and is conjugated to a fluorescent label,
- (ii) an antibody that binds to CD45 and is conjugated to a fluorescent label, and
- (iii) a nuclear stain;
- and
- (d) isolating a fetal cell using fluorescence activated cell sorting (FACS) or DEPArray, wherein the fetal cell is positive for the CK and nuclear stain and negative for CD45;
- wherein the fetal cell is a fetal nucleated red blood cell (fnRBC) or a trophoblast.
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A method for isolating fetal cells from a sample from a pregnant subject, comprising:
- (a) contacting the sample comprising an anti-epidermal growth factor receptor (EGFR) antibody and an anti-CD105 antibody;
- (b) isolating cells bound to the anti-EGFR antibody and/or anti-CD105 antibody, thereby producing an enriched sample;
- (c) contacting the enriched sample with a first labeled antibody that binds to a fetal cell marker;
- and
- (d) isolating a fetal cell that is bound to the first labeled antibody;
- wherein the fetal cell is a fetal nucleated red blood cell (fnRBC) or a trophoblast.
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Dependencies are inferred from the claim text.