Methods for purifying biological cells
This invention relates to devices and methods for purifying biological cells. For example, viable tumor, stem, immune and sperm cells can be purified from a complex biological sample using a pipette tip column. Methods of the invention can aid research, diagnosis and treatment of cancer.
1. A method for purifying viable cells, comprised of:
a) providing at least one fritted column, wherein the fritted column is comprised of a packed bed of medium, wherein the packed bed of medium has a bed volume, wherein the packed bed of medium is retained between two frits, wherein the packed bed of medium is comprised of particles, wherein the particles are comprised of an affinity group, wherein the affinity group is an antibody or a FAB;
b) providing a biological sample, wherein the sample is comprised of viable cells, wherein the viable cells are not labeled with antibodies, 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 fritted column, wherein the sample is aspirated through the packed bed of medium;
d) expelling the biological sample by reversing the flow direction, wherein the biological sample is expelled through the packed bed of medium;
e) repeating steps (c) and (d) multiple times, wherein the biological sample is not incubated on the fritted column, whereby a portion of the viable cells are captured by the affinity groups on the particles in the packed bed of medium;
f) passing a wash solution through the fritted column; and
g) eluting the cells by passing an eluent through the fritted column, wherein the eluted cells are intact and viable, wherein the eluted cells maintain their integrity, and wherein the method is at least partially performed by a timed computer-controlled program.
2. The method of claim 1 , wherein the fritted column is a pipette tip.
3. The method of claim 1 , wherein the sample is comprised of biological fluids or tissues.
4. The method of claim 1 , wherein the sample is comprised of bacterial cells, animal cells or plant cells.
5. The method of claim 1 , wherein following step (f) the captured cells are labeled on the fritted column.
6. The method of claim 1 , wherein the antibodies are covalently attached to the particles with a linker.
7. The method of claim 1 , wherein the antibodies are tagged with a drug.
8. The method of claim 1 , wherein the particles are comprised of protein A or protein G.
9. The method of claim 1 , wherein the method is used for negative selection and viable cells captured by the antibody affinity groups are not the cells of interest.
10. The method of claim 1 , wherein the eluted cells are selected from the group consisting of tumor cells, circulating tumor cells, immune cells, sperm cells, white blood cells, cancer cells, E. coli cells, T cells, B cells and stem cells.
11. A method for purifying viable cells, comprised of:
a) providing at least one fritted column, wherein the fritted column is comprised of a packed bed of medium, wherein the packed bed of medium has a bed volume, wherein the packed bed of medium is retained between two frits, wherein the packed bed of medium is comprised of particles, wherein the particles are comprised of an affinity group, wherein the affinity group is an antibody or a FAB;
b) providing a biological sample, wherein the sample is comprised of viable cells, wherein the viable cells are not labeled with antibodies, 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 fritted column, wherein the sample is aspirated through the packed bed of medium;
d) expelling the biological sample by reversing the flow direction, wherein the biological sample is expelled through the packed bed of medium;
e) repeating steps (c) and (d) multiple times, wherein the biological sample is not incubated on the fritted column, whereby a portion of the viable cells are captured by the affinity groups on the particles in the packed bed of medium;
f) passing a wash solution through the fritted column; and
g) eluting the cells by passing an eluent through the fritted column, wherein the eluted cells are intact and viable, wherein the eluted cells maintain their integrity, wherein the method is at least partially performed by a timed computer-controlled program, and wherein the entire method is performed in less than one hour.