Purification of humanized monoclonal antibodies by membrane-based hybrid bioseparation technique
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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.
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