The expansion of hybridoma cell lines, once ELISA-based data has confirmed specificity and general affinity of an antibody to its antigen, will proceed in culture using media supplemented by fetal bovine serum (FBS). While acting as an invaluable growth supplement, the wide range of proteins found in FBS,
and some at relatively abundant levels, will prove problematic for purification of a monoclonal antibody(mAb).
ProMab makes every effort to ensure when using our in vitro monoclonal antibody service, you will receive a monoclonal antibody (mAb) of highest purity. Therefore, once the expansion and Antibody production requires 2 – 4 weeks and ranges from $800 to $1,100 with purification establishment of a hybridoma is complete, cells will be gradually weaned away from media containing FBS to a completely serum-free media. As soon as the serum-free media growth characteristics of the hybridoma line approximate those of cells in the presence of FBS, cell growth will continue either in shake-flasks, or in spinner flasks (while monoclonal antibodies, mAb, yield would often dictate a preference for the use of spinner flasks, growth properties of specific hybridoma’s will require the use of the shake flask exclusively). As cells continue to grow, media will be withdrawn and replaced continuously until a minimum volume of 1 liter of antibody-containing media has been collected.
Serum-free media containing the monoclonal antibody (mAb) is then passed over a Protein G column, thereby binding, and purifying the monoclonal antibody (mAb) from all other media components.