Purification of humanized monoclonal antibodies by membrane-based hybrid bioseparation technique.
We describe a rapid, inexpensive and scalable hybrid bioseparation technique for one-step purification of humanized monoclonal antibodies from mammalian cell culture supernatant. It involves the selective and reversible retention of the monoclonal antibody within a membrane module utilizing a combination of microfiltration of precipitated antibody and simultaneous hydrophobic interaction-based membrane chromatography of soluble antibody on the same microfiltration membrane. The retained monoclonal antibody is subsequently recovered from the membrane module in a highly pure form by changing the solution condition to that which favours simultaneous antibody dissolution and elution from the membrane. This combination of separation principles results in high-capacity, high-resolution separation. The monoclonal antibody purities and recoveries obtained in the experiments described in this paper were very high.
Wang
Lu
L
Department of Chemical Engineering, McMaster University, 1280 Main Street West, Hamilton, Ontario, Canada L8S 4L7.
Kanani
Dharmesh M
DM
Ghosh
Raja
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D016466:CHO Cells; D002678:Chimera; D002846:Chromatography, Affinity / Q000379:methods*; D006224:Cricetinae; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007158:Immunologic Techniques
D000911:Antibodies, Monoclonal; D003470:Culture Media