Centrifugation.

• Centrifugation coupled with depth filtration has been employed for the initial recovery of
therapeutic proteins for years and is well suited for pilot and commercial scale
manufacturing.
• Disk-stack continuous centrifuges are capable of removing cells and large cell debris
• Hermetic centrifuges are used harvest the mammalian cell  
Depth filtration.
Depth filters are traditionally used in the clarification of cell culture broths, to maintain
capacity on membrane filters or to protect chromatography columns or virus filters.
Depth filters are typically composed of cellulose, a porous filter-aid such as diatomaceous
earth and an ionic charged resin binder.
The pore size range of up to 10 µm and removes whole cells and large particles
The pore size 0.2 µm remove a wide range of process contaminants and impurities
Ammonium sulphate precipitation
 This method is used to isolate antibodies from serum, cell culture supernatant, or ascites
fluid.
 Different proteins, including antibodies, precipitate at specific concentrations of ammonium
sulphate. For example, antibodies precipitate at 40-50% ammonium sulphate.

Physicochemical fractionation
 Physicochemical fractionation describes methods that separate antibodies by size, charge, or
chemical properties.
 The different classes of immunoglobulin (Ig), such as IgM and IgG, often have similar
amino acid compositions, solubility, and structure.
Overall schematic flowchart of monoclonal antibody purification
Questions?