Monoclonal Antibody Production


Hybridoma lines are generally created by the HAT (hypoxanthine-aminopterin-thymidine) selection method. These aim to select out unfused spleen/ myeloma cells to result in a culture enriched in hybridoma cells. Hybridomas can produce monoclonal antibodies (mAbs) with high titre only if induced under high-stress situation. There is no generalized and optimized cell culture protocol for hybridoma cultures because optimum growth conditions for every individual clone will vary.

A 2D system, e.g. T-175 flasks, or any flask-based cell culture system, is initially used to grow and expand the cells using cell-growth optimal media followed by cell-induction medium. This technique can be applied in combination with partial serum starvation to acclimatise the hybridoma cells.

The purpose of this application note is to provide scientists with a generic condition for the first CelCradle optimization experiment. Key components and the design methodology deployed in single-use CelCradle-500AP bottle in perfusion mode will be highlighted. It is recommended for one to read this application note prior to conducting the first experimental run.