Methods and devices from isolation of tumor cells
Described herein are methods and compositions for isolating cancer cells using positively charged surfaces, such as beads, microparticles, and capillary tubes. The methods and compositions for isolation can be used in cancer diagnosis, treatment, and therapeutic preparation.
1 . A method of removing cancer cells from a biological sample, comprising:
a) passing a biological sample comprising cancer cells obtained from a subject over an isolation matrix comprising a positively charged surface, wherein the positively charged surface comprises glass that is functionalized with a positively charged functional group selected from the group consisting of an amine, an aldehyde, polyethylenimine (PEI), a guanidine group, and a combination thereof; and
b) collecting the biological sample that flows through the isolation matrix comprising the positively charged surface in a first aliquot,
wherein the cancer cells bind to the isolation matrix.
2 . The method of claim 1 , wherein the positively charged surface is selected from a bead, a microparticle, a capillary tube, a blood collection tube, a microscope slide, and a microscope slide coverslip.
3 . The method of claim 1 , wherein the biological sample comprises blood.
4 . The method of claim 1 , further comprising lysing the cancer cells bound to the isolation matrix, thereby obtaining a cancer cell lysate.
5 . The method of claim 1 , wherein the first aliquot is administered back into the subject.
6 . The method of claim 1 , further comprising detecting the presence of the cancer cells in the biological sample via cell staining or cell lysis followed by PCR amplification.
7 . The method of claim 1 , wherein the positively charged surface is a glass bead functionalized with an amine, an aldehyde, PEI, or a guanidine group.
8 . The method of claim 7 , wherein the positively charged surface is a glass bead functionalized with an amine.
9 . The method of claim 7 , wherein the positively charged surface is a glass bead functionalized with an aldehyde.
10 . The method of claim 7 , wherein the positively charged surface is a glass bead functionalized with PEI.
11 . The method of claim 7 , wherein the positively charged surface is a glass bead functionalized with a guanidine group.
12 . A method of detecting a cancer cell in a patient, comprising:
a) passing a biological sample from the patient through a functionalized glass capillary tube or an isolation matrix of functionalized glass microspheres, wherein the functionalized glass capillary tube or the functionalized glass microspheres comprise amine, polyethylenimine (PEI), and/or guanidine groups that are attached to the glass capillary tube or glass microsphere via a siloxane bond, and wherein the functionalized glass capillary tube or the functionalized glass microspheres bind cancer cells;
b) eluting the biological sample that flows through the functionalized glass capillary tube or the isolation matrix; and
c) detecting the presence of a cancer cell bound to the functionalized glass capillary tube or functionalized glass microspheres.
13 . The method of claim 12 , wherein the patient has a hematological cancer or a malignant cancer.
14 . The method of claim 12 , wherein the biological sample is passed through the isolation matrix of functionalized glass microspheres.
15 . A method of binding a cancer cell from a biological sample, comprising:
a) contacting a biological sample comprising a cancer cell with an isolation matrix comprising glass beads having surfaces that are functionalized with a positively charged functional group which may be any one or more of an amine, an aldehyde, polyethyleneimine (PEI), or a guanidine group;
b) washing the isolation matrix to remove the biological sample from the isolation matrix, wherein the cancer cell binds to the isolation matrix;
c) lysing the cancer cell, thereby providing a lysate; and
d) detecting in the lysate one or more nucleic acids.
16 . The method of claim 15 , wherein the biological sample is obtained from a subject suspected of having a cancer.
17 . The method of claim 16 , wherein the cancer is a hematological cancer or comprises a solid tumor.
18 . The method of claim 15 , wherein detecting in the lysate one or more nucleic acids comprises performing RT-PCR on mRNA isolated from the lysate.