Antibody Production from a Batch Overgrowth Culture of Transfected Mammalian Cells

Seed cells at 2 x 10⁵ cells per milliliter in 100 milliliters of serum-free medium in two flasks. Incubate the cells at 37 degrees Celsius and 5% carbon dioxide at 120 rotations per minute for 24 hours. To the nutrient-supplemented culture, add tryptone to a final concentration of 0.5%. Then, use serum-free medium to equalize the volume of the unsupplemented culture. Incubate the cells for an additional seven days.

From the eighth day of culturing forward, count the cells daily using a hemocytometer to monitor cell viability. Once the cell viability reaches less than 80%, harvest the culture by centrifugation at 3,000 times g for 15 minutes. Then, filter-sterilize the supernatant through a 0.22-micrometer filter. Store the supernatants at 4 degrees Celsius in the short term or freeze at degrees Celsius long-term.