Method of isolating circulating tumor cells

Patent metadata

Field	Value
Publication number	US-10161939-B2
Application number	US-201314765071-A
Country	US
Kind code	B2
Filing date	May 31, 2013
Priority date	Feb 2, 2013
Publication date	Dec 25, 2018
Grant date	Dec 25, 2018

Abstract

Provided are methods for detecting or isolating circulating tumor cells (CTCs) in a subject. The methods may include detecting the expression of at least one epithelial mesenchymal transition (EMT) biomarker. Further provided are kits for detecting or isolating CTCs. The kits may include antibodies to at least one EMT biomarker. Further provided are methods of predicting the responsiveness of a subject to a cancer drug, methods of targeting delivery of a cancer drug in a subject, methods of providing a cancer prognosis to a subject, and methods for following the progress of cancer in a subject.

First claim

What is claimed is:

1. A method for isolating, capturing, or enriching a circulating tumor cell from a patient, the method comprising:
a) obtaining a biological sample from a patient;
b) obtaining at least one capture binding protein, wherein the capture binding protein is linked to a solid phase to form a solid phase-capture binding protein complex, and wherein the capture binding protein is an epithelial-mesenchymal transition (EMT) biomarker capture antibody;
c) contacting the biological sample with the solid phase-capture binding protein complex for a time sufficient to allow the solid phase-capture binding protein complex to bind at least one EMT biomarker on the circulating tumor cell to form a solid phase-capture binding protein-circulating tumor cell complex, wherein the EMT biomarker is at least one of OB-cadherin, N-cadherin, vimentin, E-cadherin, FGFR2 splice variant isoforms, or CD133;
d) separating the solid phase-capture binding protein-circulating tumor cell complex from the sample and unbound magnetic particle-capture binding protein complexes by application of an external magnetic field on the sample, thereby isolating, capturing, or enriching the circulating tumor cell; and
e) confirming the circulating tumor cell, wherein confirming comprises β-catenin expression detection and CD31 expression detection, wherein the circulating tumor cell is confirmed if β-catenin expression is positive and CD31 expression is negative.

2. The method of claim 1 , wherein confirming the circulating tumor cell further comprises at least one of DAPI staining and CD45 detection.

3. The method of claim 2 , wherein the circulating tumor cell is further confirmed if DAPI staining is positive and CD45 expression is negative.

4. The method of claim 1 , wherein the circulating tumor cell has a mesenchymal phenotype.

5. The method of claim 1 , wherein the patient has cancer.

6. The method of claim 1 , further comprising determining the presence or absence of at least one prostate cancer-specific genomic event.

7. The method of claim 6 , wherein the at least one prostate cancer-specific genomic event is selected from the group consisting of androgen receptor amplification, phosphatase and tensin homolog (PTEN) loss, gene fusion of transmembrane protease, serine 2 (TMPRSS2) gene and ETS related (ERG) gene, and combinations thereof.

8. The method of claim 1 , wherein the biological sample comprises a tissue sample or a fluid sample from an organism.

9. The method of claim 1 , wherein the β-catenin expression detection comprises contacting the solid phase-capture binding protein-circulating tumor cell complex with a first staining reagent that binds β-catenin, and wherein the CD31 expression detection comprises contacting the solid phase-capture binding protein-circulating tumor cell complex with a second staining reagent that binds CD31.

10. The method of claim 9 , wherein the first staining reagent comprises an antibody that binds β-catenin.

11. The method of claim 9 , wherein the second staining reagent comprises an antibody that binds CD31.

Assignees

• Univ Duke
• Janssen Diagnostics Llc

Inventors

• Rao Galla Chandra
• Connelly Mark C
• Garcia-Blanco Mariano A
• Armstrong Andrew J
• Bitting Rhonda L

Classifications

• G01N33/57555

  of the prostate · CPC title

• G01N33/57515

  of the breast · CPC title

• G01N33/5759

  involving compounds localised on the membrane of tumour or cancer cells · CPC title

• G01N33/5758 Primary

  involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites · CPC title

• G01N33/575

  for cancer · CPC title