Mol. Biotechnol., 20, 29-40 (2002)
Cunha, M. T., Aires-Barros, M. R.
The production of foreign proteins using selected host with the necessary
posttranslational modifications is one of the key successes in modern biotechnology. This
methodology allows the industrial production of proteins that otherwise are produced in
small quantities. However, the separation and purification of these proteins from the
fermentation media constitutes a major bottleneck for the widespread commercialization of
recombinant proteins. The major production costs (5090%) for typical biological
product resides in the purification strategy. There is a need for efficient, effective,
and economic large-scale bioseparation techniques, to achieve high purity and high
recovery, while maintaining the biological activity of the molecule. Aqueous two-phase
systems (ATPS) allow process integration as simultaneously separation and concentration of
the target protein is achieved, with posterior removal and recycle of the polymer. The
ease of scaleup combined with the high partition coefficients obtained allow its potential
application in large-scale downstream processing of proteins produced by fermentation.
The equipment and the methodology for aqueous two-phase extraction of proteins on a large scale using mixer-settler and column contractors are described. The operation of the columns, either stagewise or differential, are summarized. A brief description of the methods used to account for mass transfer coefficients, hydrodynamics parameters of hold-up, drop size, and velocity, back mixing in the phases, and flooding performance, required for column design, is also provided.
Keywords: protein extraction; aqueous two-phase systems
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