RAb M5114 Production results from the Fibercell Hollow Fiber System. Cartridge used was the C2011 20 kD MWCO polysulfone fiber and medium used was DMEM with 10% CDM-HD. Protocol followed was per the FiberCell Systems manual. In the gel shown, purified M5114 was a positive control purified from Protein G column, lane 1. Lanes 2,3, and 4 are harvests from the cartridge on day 7, 9 and 12. No purification has been performed. Note how clean the harvests are and free from extraneous proteins. Lane 5 is 10X concentrated supernatant from a T-flask using DMEM and 10% CDM-HD. Lane 6 is from a T flask using DMEM and 10% serum. Based on the gel, it is estimated that the concentration of supernatant from HFB was about 2.5 mg/mL. 140 mL of supernatant was harvested to date for a total about 350 mg of non-purified antibody. Both SDS-PAGE gel and gel filtration analysis showed that the antibody from the hollow fiber bioreactor was very clean and properly folded. Data courtesy of Liying Lu, University of Massachusetts, Worcester, Worcester, Massachusetts.
