Method for selecting high producing cell lines
The invention provides methods for the rapid identification and selection of cell lines suitable for biopharmaceuticals production, which do no utilize animal derived components.
1. A method for selecting one or more mammalian cells expressing secreted antibody for use as a biopharmaceutical, the method comprising the steps of:
(a) providing a population of mammalian cells in a medium free from animal-derived components, wherein the cell population comprises at least one cell expressing said secreted antibody;
(b) contacting the cell population with a label which directly binds to said antibody on the surface of the cell;
(c) detecting said label bound to the antibody on the surface of said cell; and
(d) selecting said cell based on the presence of said label bound to the antibody on the surface of said cell;
wherein the label is selected from the group consisting of Protein A, Protein G or Protein L; and whereby one or more mammalian cells expressing a secreted antibody for use as a biopharmaceutical is selected.
2. The method according to claim 1 , wherein said Protein A, Protein G or Protein L is fluorescent.
3. The method according to claim 1 , wherein the detecting step comprises detecting a relative amount of said label bound to the surface of said cell.
4. The method according to claim 1 , wherein the selecting step comprises selecting the cell or cells from the population which show the highest amount of label bound thereto.
5. The method according to claim 1 , further comprising the step of:
(e) isolating said selected cell or cells from the population of cells.
6. The method according to claim 1 , wherein the secreted antibody is a domain antibody.
7. The method according to claim 1 , wherein the mammalian cell is selected from the group consisting of CHO, CHO K1, CHO DG44, NSO, COS-1, COS-7, H EK293, H K21, PerC6, H EK293, 293T, Vero, AGE1.CR, HT1080, TE671, Namalwa, and SP2/0.
8. The method according to claim 1 , wherein the steps of detecting, selecting and/or isolating said cell is carried out by fluorescence activated cell sorting.
9. The method according to claim 8 , wherein said label is fluorescent Protein A, Protein G, or Protein L; wherein the step of detection comprises determining a relative fluorescence of said label bound to the antibody on the surface of the cell, wherein a relatively higher fluorescence is indicative of a higher relative expression of said antibody; and wherein selection step comprises selecting the cell or cells from the population that have a relatively higher level of fluorescence.