Continuous Isolation of rFVIII from Mammalian Cell Culture

Abstract

This is a proof-of-principle study on the potential application of continuous annular chromatography for the direct capture of protein product from perfusion culture supernatant, and the first step in its evaluation as potential platform technology for large scale production of recombinant protein drugs. A standard P-CAC column was specifically modified for isolation operation. Peak wobbling due to inhomogeneous flow was identified as a performance limiting factor. As a real world application, the continuous isolation of recombinant Factor VIII from culture supernatant using ion exchangeresin is presented. A sterile P-CAC prototype has been developed to allow on-line coupling to continuous perfusion fermenters.