Devices and methods for purification, detection and use of biological cells
This invention relates to devices and methods for purifying, detecting and using biological cells. A variety of cell types including viable tumor, stem, immune and sperm cells can be purified from a complex biological sample using a column, including a pipette tip column. Methods of the invention can aid research, diagnosis and treatment of cancer. Purified viable cells can be detected on the column or eluted from the column and detected. Cells on a column can be used as a stationary phase for liquid chromatography. Cells may be removed, recovered and analyzed.
1. A method of making a cell-based chromatography column and performing chromatography, comprising the steps of:
a) providing at least one column, wherein the column is comprised of a packed bed of particles, wherein the packed bed of particles has a bed volume, wherein the packed bed of particles is retained between two frits, a lower frit and an upper frit, wherein the packed bed of particles is comprised of physical channels, and wherein the particles are capable of capturing cells;
b) providing a biological sample comprised of viable cells, wherein the biological sample has a sample volume, wherein the sample volume is larger than the bed volume;
c) aspirating the biological sample through the column using bidirectional flow, wherein the sample is aspirated through the lower frit, into the packed bed of particles and then through the upper frit;
d) expelling the biological sample, wherein the biological sample is expelled through the upper frit, into the packed bed of particles and then through the lower frit;
e) repeating steps (c) and (d) multiple times, wherein the viable cells pass through the physical channels in the packed bed of particles without being trapped, wherein a portion of the viable cells are captured on the particles, whereby the viable cells captured on the column comprise the cell-based chromatography column;
f) performing chromatography by providing a second sample, wherein the second sample is comprised of antibodies, wherein the antibodies are further comprised of a drug or drug candidate, wherein the drug or drug candidate is specific for a cell surface marker present on diseased cells; and
g) passing the second sample through the cell-based chromatography column, wherein the second sample is passed through the chromatograph using a chromatography method selected from the group consisting of step gradient chromatography, displacement chromatography, partitioning chromatography and breakthrough curve chromatography.
2. The method of claim 1 , wherein step (g) is performed using unidirectional flow.
3. The method of claim 1 , wherein step (g) is performed using bidirectional flow.
4. The method of claim 1 , wherein following step (g), the cells are removed from the column and analyzed.
5. The method of claim 1 , wherein the particles in the packed bed are impervious to reagents.
6. The method of claim 1 , wherein the method is automated.
7. A method of making a cell-based chromatography column and performing chromatography, comprising the steps of:
a) providing at least one column, wherein the column is comprised of a bed of particles, wherein the bed of particles is retained between two frits, a lower frit and an upper frit, wherein the bed of particles is comprised of physical channels, and wherein the particles are capable of capturing cells;
b) providing a biological sample comprised of viable cells, wherein the viable cells are comprised of a surface marker;
c) aspirating the biological sample into the column through the lower frit;
d) expelling the biological sample out of the column through the lower frit;
e) repeating steps (c) and (d) multiple times, wherein the viable cells pass through the physical channels in the bed of particles without being trapped, wherein a portion of the viable cells are captured on the particles in the bed of particles, wherein the viable cells are attached to the particles via the surface marker, whereby the viable cells captured on the column comprise the cell-based chromatography column;
f) performing chromatography by providing a second sample, wherein the second sample is comprised of antibodies, wherein the antibodies are further comprised of a drug or drug candidate, wherein the drug or drug candidate is specific for a cell surface marker present on diseased cells; and
g) passing the second sample through the cell-based chromatography column, wherein the second sample is passed through the cell-based chromatography column using bidirectional flow.
8. The method of claim 7 , wherein the particles impervious to reagents.
9. The method of claim 7 , wherein the method is automated.
10. The method of claim 1 , wherein the viable cells are cancer cells.
11. The method of claim 7 , wherein the viable cells are cancer cells.
12. The method of claim 1 , wherein the antibody drug or antibody drug candidate is specific for infectious disease, diabetes, heart disease, Parkinson's, Alzheimer's or liver disease.
13. The method of claim 7 , wherein the antibody drug or antibody drug candidate is specific for infectious disease, diabetes, heart disease, Parkinson's, Alzheimer's or liver disease.