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Devices and methods of cell capture and analysis

Inactive Publication Date: 2012-04-26
BIOCEPT INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0009]In one aspect, the invention provides a method for capturing biological targets from solution. In this aspect, the present invention is based, in part, on the discovery that pre-labeling

Problems solved by technology

These current approaches suffer from several technical difficulties, one of which is the problem of non-specific binding.
In addition, the subsequent in situ analysis of cells on the channel by staining and hybridization procedures may subject the cells to harsh and denaturing conditions.
These washing and analysis procedures can compromise the initial capture of the desired molecule or cell by subjecting the binding partner to conditions that may cause the binding partner to degrade, lose some of its conformational structure, or become detached from the solid support.
Further still, existing methods for analyzing circulating cells (e.g., as captured from a patient sample) for malignancy, such as staining cells for cytokeratin (CK), have limitations as markers for identifying and / or evaluating circulating tumor cells.

Method used

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  • Devices and methods of cell capture and analysis
  • Devices and methods of cell capture and analysis
  • Devices and methods of cell capture and analysis

Examples

Experimental program
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example 1

Construction of Basic Micro-Channel Device

[0114]One embodiment of a micro-channel device for separating biomolecules or cells is shown in FIG. 1. The device comprises a substrate or support 11 which is formed with a flow path that includes a micro-channel 13 to which sample liquid is to be supplied through an opening or well 15 that serves as an entrance or inlet at a first end of the device and an opening 19 that serves as an outlet at the second end of the device. The cross-section of the collection region 17 is greater than that of an inlet section 18 that leads there into from the inlet opening 15. The inlet section contains one or more pairs of axially aligned divider / supports 21 just upstream of where it widens at the end of the region 18 to enter the collection region 17. These central dividers break the flow into two or more paths and serve to distribute the flow of liquid more evenly as it is delivered to the entrance end of the collection region 17. The collection region c...

example 2

Comparison of Cell Capture Rates Between Pre-Labeled Micro-Channels and Pre-Labeled Cells

[0118]As described in U.S. Published Application No. 2006 / 0160243, filed Jan. 18, 2005 and elsewhere (Nagrath et al. (2007) Nature, Vol. 450(7173):1235-9), previous devices for capturing cells of interest comprised a micro-channel that was derivatized with an antibody that was specific to antigens on the cells of interest. The suspension containing the rare cells of interest was then passed through the channel and cells were captured by the cell-specific antibody (FIG. 2).

[0119]While the level of antigen expression can be determined in cultured cells and on clinical tissue samples such as tumors, it is not known precisely how many antigens are available on the surface of a circulating tumor cell (CTC). It is known that tumors are highly heterogeneous and that cells detached from the tumor into the blood can change their expression levels of antigen. Therefore, it is most likely that CTCs are a h...

example 3

Use of Multiple Antibodies Increases the Capture Rate of Target Cells

[0127]It has traditionally been considered most efficient to pre-load an antibody onto the channel. However, the negative effects on cell capture of loading a channel with multiple antibodies have not been previously considered. An advantage of using a micro-channel coated with a general binding partner (e.g. antibody or protein) of an antigen-specific antibody is that multiple antibodies can be added to a cell suspension to pre-label cells without lessening the availability of any single antibody. Because multiple antigen sites on a cell are not mutually exclusive, when adding multiple antibodies to the cell suspension the capture efficiency on the channel is not diminished for any single antibody. By way of example, if the channel could accommodate 100 antibody sites and a mixture of 5 different antibodies were added to coat the channel, then each antibody would occupy ˜20% of the channel space. Thus, the potenti...

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Abstract

The present invention provides a device for isolating target biomolecules or cells from samples, particularly biological samples. In particular, the device comprises a loading mixture, which contains the biological sample and a first binding entity that specifically binds to the target biomolecule or target cell; and a micro-channel coated with a second binding entity that binds directly or indirectly to the first binding entity. Methods of capturing, detecting, and / or evaluating target biomolecules or target cells (e.g. cancer cells) in biological samples are also disclosed.

Description

RELATED APPLICATIONS[0001]This application is a continuation-in-part of U.S. application Ser. No. 12 / 730,738 filed Mar. 24, 2010; which is incorporated herein by reference in its entirety. The present application also claims priority to U.S. Provisional Application No. 61 / 431,385, filed Jan. 10, 2011; PCT Application No. PCT / US2010 / 028499 filed Mar. 24, 2010; U.S. Provisional Application No. 61 / 235,615 filed Aug. 20, 2009; U.S. Provisional Application No. 61 / 163,009, filed Mar. 24, 2009, and U.S. Provisional Application No. 61 / 298,871, filed Jan. 27, 2010; each of which are hereby incorporated by reference in their entireties.FIELD OF THE INVENTION[0002]The present invention relates to micro-channel devices for capturing targets, such as cells and molecules of interest from solutions, as well as to post-capture analysis of circulating cells. In certain embodiments, the present invention relates to methods and devices for capturing target cells (e.g. circulating tumor cells) from phy...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N21/64C12M1/34C12Q1/68G01N33/574G01N33/82
CPCG01N33/54306G01N33/56966C12Q1/6886G01N2333/4742G01N33/57496G01N33/57446G01N2333/71G01N33/57434G01N33/57423G01N33/57415C12Q2600/156G01N33/57492
Inventor MIKOLAJCZYK, STEPHEN D.PIRCHER, TONYBISCHOFF, FARIDEH Z.TSINBERG, PAVEL
Owner BIOCEPT INC
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