Ascites Production Process

 

The ascites method is one of the quickest for producing large amounts of monoclonal antibodies. One mouse can produce 3-5 ml of ascites fluid after undergoing several extraction cycles after inoculation. Inoculating multiple mice simultaneously can produce large batches of a monoclonal antibody.

Priming the Mouse

The mouse’s peritoneal cavity needs to be primed before hybridoma cells are injected. This promotes growth and antibody production in the hybridoma cells. Pristine is often used, it induces an immune reaction and prevents fluid from leaking out of the peritoneum. The mouse is then primed for two weeks prior to hybridoma cells are injection.

Injecting the Hybridoma Cells

Hybridoma cells are grown in tissue culture prior to injection into a mouse for ascites antibody production. The process is timed such that the cells are in the growth phase in tissue culture at the time during which they are injected. It is also important that the hybridoma be in the same genetic background as the host mouse to avoid host rejection. The number of cells that are injected ranges from hundreds of thousands to millions; with millions being on the high end of this procedure.

Incubation Times Before First Tap

Ascites fluid is allowed to accumulate for about one to two weeks before any is taken out. Care is taken to observe the condition of the mice daily. The ascites fluid is removed from the peritoneal cavity, a process referred to as “tapping.”

Isolation of Aqueous Ascites Component

After the ascites fluid is extracted from the mouse, it is spun down in a centrifuge. This process separates the lipid compartment from the cellular compartment to the bottom. In the middle is the aqueous compartment that is referred to as ascites fluid, and that contains the desired antibodies.