Case Study, Hybridomas Cultured with CDM-HD

Two hybridomas were cultured in separate bioreactors using High Glucose DMEM + 10% CDM media + antibiotics. The first week is the setup of the apparatus and the seeding of the system.  Weeks 2 through 4 are harvest weeks (we only do production for a month).  The supernatants collected from the bioreactor were dialyzed (10kDa MWCO) against 2 x 1L changes of PBS with 0.3M EDTA, and then against 2 X 1L changes of PBS. Protein concentration was determined by BCA (Pierce) and purity checked on a 12% SDS-PAGE gel. Note, there is no antibody purification step included.

MAB 1 was growing in Invitrogens serum free media before seeding into the bioreactor. 

Mab 1 – 2.6mg/ml week two and three harvest (40 ml) Yield 104 mg
Mab 1 – 3.2mg/ml week four harvest (20 ml) Yield 64 mg

Three week yield 168 mg in collected in 60mls of media. Total media consumed 9 L DMEM + 10% CDM.  Average antibody concentration 2.8 mg/ml.

Mab 2 was growing in RMPI + 20% FBS before seeding into the bioreactor. It took a few days for the cells to adapt to DMEM + 10% CDM HD but no specific adaptation protocol was followed.

Mab 2 – 0.8 mg/ml week two harvest (25ml) Yield 20 mg
Mab 2 – 3.2 mg/ml week three harvest (20ml) Yield 64 mg
Mab 2 – 3.0 mg/ml week four harvest (25ml) Yield 75 mg

Three week yield 159 mg in collected in 70 ml of media.  Total media consumed 11 L DMEM +10% CDM.  Average antibody concentration 2.3 mg/ml.

Note how clean the antibody in the gel is. There are no exogenous proteins.