Methods and kits for the detection of circulating tumor cells in pancreatic patients using polyspecific capture and cocktail detection reagents

Abstract

A highly sensitive assay is disclosed which combines immunomagnetic enrichment with multiparameter flow cytometric or image cytometry to detect, enumerate and characterize carcinoma cells in the blood. The present invention incorporates the conjugation of different antibodies to the same ferrofluid. This has the effect of making the ferrofluid polyspecific with respect to the antigens that the ferrofluid will bind. The multiple antibodies present on the same ferrofluid do not appear to block or otherwise interfere with each other. Such ferrofluids have the highly desirable effect of being able to bind specifically to more than one type of cell. The assay is especially useful to enable the capture of CTCs that have low EpCAM expression, but high expression of other tumor markers; Accordingly, the assay facilitates the biological characterization and staging of carcinoma cells.

Description

[0001] Related Patent Applications [0002] This patent application claims the benefit of priority to US Provisional Application No. 61 / 393,036, filed October 14, 2010, which is hereby incorporated by reference in its entirety. technical field [0003] The present invention relates to the fields of oncology and diagnostic testing. The invention is useful for cancer screening, staging, monitoring chemotherapy treatment response, cancer recurrence, and the like. More specifically, the present invention provides reagents, methods and test kits that facilitate the analysis and enumeration of tumor cells or other rare cells isolated from biological samples. Background technique [0004] Enumeration of circulating tumor cells (CTCs) in patients with metastatic breast, prostate and colon cancer using the CellSearch CTC assay predicts patient survival and enables monitoring of treatment response. In addition, CTCs can be characterized for a variety of molecular markers that have b...

AI Technical Summary

Problems solved by technology

However, there is a limit regarding the number of magnetic particles that can be added, since rare cells mixed with the bulk of magnetic particles after separation can be replaced by rare cells mixed with other blood cells

The latter condition does not significantly improve the ability to count or examine cells of interest

[0021] There is another disadvantage of using large particles to isolate cells at rare frequencies (1 to 50 cells / ml blood)

Despite the fact that large magnetic particles allow the use of external gradients of very simple design and relatively low magnetic gradients, large particles tend to cluster around cells in a cage-like formation, making cells difficult to visualize or analyze

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