Game-changing innovations
Although process redesigns and traditional technologies can contribute to the development of downstream processes, they provide
only incremental improvements that marginally increase process efficiency. Incremental or evolutionary technologies have been
the mainstay of the bioprocessing industry for the past 20 years, and column chromatography provides one of the best examples
of this phenomenon in action (29). However, these slow marginal gains are already beginning to decline, and it is becoming
difficult to envisage how sustainable processing can continue without a major injection of downstream-processing capacity.
One way to address this concern is to embrace genuinely novel technological approaches that change the rules of the game.
The fringes of the biopharmaceutical industry are populated by companies that survive on innovation, and some of these innovations
are disruptive in the sense that their influence on the industry is unpredictable and could contribute to a radical change
in bioprocessing.
Most technological innovations in bioprocessing have been incremental, but there are several recent examples of disruptive
innovations that have challenged the established business model and caused real grass-roots change in the industry. Again,
many of these changes have affected upstream productivity first (e.g., disposable bioreactors and buffer/media storage bags),
but we are also seeing examples in downstream processing (e.g., the introduction of simulated moving bed chromatography, expanded
bed chromatography, monoliths, and membrane adsorbers) (1). These innovations have taken hold in niche markets but are now
beginning to adopt mainstream positions. Disposable modules for downstream processing occupy a more mature status in the development
cycle (30). The use of disposable filter modules is now an industry standard and where filters first left their footprint,
membrane adsorbers are set to follow (31).
Table I: Broader polishing operation window with salt-tolerant membrane chromatography.
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Membrane technology.
Disposable membranes adsorbers are beginning to replace traditional chromatography in a number of settings, just as disposable
membrane filters replaced steel mesh filters. Indeed, membrane filters have evolved into charged filtration devices that use
the principles of both sieving and chemical selection to improve filtration performance, thereby creating a precedent for
the use of membrane chromatography in downstream processing. After a period of inertia, the benefits of membrane chromatography
are now fairly well established, and manufacturers are willing to consider them as a genuine alternative to fixed columns
rather than a step in the dark (32, 33). In contrast to resin-based flow through processes, membrane chromatography involves
the use of thin, synthetic microporous or macroporous membranes stacked in layers within a disposable cartridge (34). The
footprint of such devices is much smaller than columns with a similar output. A range of membranes is available with functional
groups equivalent to the corresponding resins (e.g., membranes containing activated quaternary ammonium groups for anion exchange,
or phenyl groups for hydrophobic interaction chromatography [HIC]), and a relatively new variant also allows salt tolerant
interaction chromatography (STIC) in high-salt buffers (35, 36). The availability of STIC membranes is an important and innovative
advance in biomanufacturing because even the most recent generation of membrane adsorbers fall short of some manufacturing
requirements when challenged with the high-conductivity feed streams often produced in high-titer processes. STIC ensures
more flexibility in process design and improves the clearance of host cell proteins and viruses in buffers containing high
concentrations of salt (see Table I). These new adsorbers therefore allow polishing to be carried out at higher load densities
without an interstitial dilution step after product capture, reducing process time and circumventing the need for additional
buffer preparation and holding.
Table I demonstrates that Sartobind STIC provides higher binding capacities for BSA, DNA, and model viruses compared with
a Q anion exchanger under high salt conditions (150 mM NaCl), thereby increasing the design space for polishing operating
conditions.
In a recent example described by the Italian biopharmaceutical company Philogen, membrane adsorbers were substituted for the
flow through and bind-and-elute steps for the polishing of a new monoclonal antibody fusion protein in Phase I–II clinical
development, achieving 90% recovery and 99.9% purity (37). The performance benefits of membranes provide value for the user,
but the complete elimination of cleaning and validation requirements is often cited as the major advantage because this avoids
the costs of the chemicals, personnel, and record-keeping, and more importantly avoids the inevitable process down time while
cleaning takes place. Spent modules are simply replaced with prevalidated new ones, available in a range of sizes and configurations
for maximum flexibility (32).
Figure 1: (a) Mechanistic comparison of solute transport in bead resins (left) and membrane adsorbers (right), where thicker
arrows represent bulk convection, thinner arrows represent film diffusion and curved arrows represent pore diffusion. (b)
Comparison of bed height in columns (left) and membrane adsorbers (right). Using membrane adsorbers is functionally equivalent
to shortening columns to near-zero length, resulting in a similarly small pressure drop that allows extremely high flow rates,
thereby reducing overall process times up to a 100-fold. In this example, both formats have a 1350 cm2 frontal surface; the column has a bed height of 15 cm; and the membrane adsorber has a bed height of 0.4 cm. The height to
frontal surface ratio is approximately 100 for the column and nearer to 3500 for the membrane device. (ALL FIGURES ARE COURTESY
OF THE AUTHOR)
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The performance advantage of membranes over resins reflects the transport of solutes to their binding sites mainly by convection,
while pore diffusion is minimal (see Figure 1a). Because of these hydrodynamic benefits, membrane adsorbers can operate at
much greater flow rates than columns, thereby considerably reducing buffer consumption and shortening the overall process
time by up to 100-fold. The use of membrane adsorbers can be viewed as the equivalent of shortening traditional columns to
near zero length to allow large-scale processes to run with only a small pressure drop at high flow rates. For example, polishing
with an anion exchange membrane can be conducted with a bed height of 4 mm at flow rates of more than 600 cm/h, and provide
a high frontal surface area-to-bed height ratio (see Figure 1b). Small-volume disposable membrane chromatography devices can
now handle up to 50 L /min/bar/m2. Even at these high flow rates, the membrane pores provide adequate binding capacity for large molecules such as viruses
and DNA, so they can play an important role in the overall viral clearance strategy for antibody purification (38, 39).
Figure 2: Selection guide for convective media, such as membrane adsorbers. HIC is hydrophobic interaction chromatography.
STIC is salt tolerant interaction chromatography.
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Disposable modules.
The flexibility of disposable modules is arguably the most important benefit in the context of the whole process, and this
reflects the broad industry perspective that manufacturing flexibility is now perhaps at least as important as capacity considering
the large numbers of products in clinical development. Process development can be streamlined and expedited because different
modules can be tested in various combinations to arrive quickly at the best overall set of process options, and the absence
of cleaning and validation requirements can shorten the time required to develop a finalized process by months or years. The
ability to replace each module completely also makes it easier to assemble process trains for new products in existing premises
without cross-contamination. The flexibility is most noticeable during scale-up because disposable devices are generally modular
and available in various sizes, and scaling up simply involves swapping one module for another with a higher capacity. It
is thus apparent that membrane devices can be scaled up with none of the attendant disadvantages of column resins, thereby
making the goal of polishing 100-kg batches of antibody entirely possible without oversizing.
Looking ahead
Innovations that take into account the current state of the industry as well as potential challenges and demands are likely
to be the most successful in the long term. At the same time, technologies borrowed from the edge of research always come
with risks that must be evaluated by manufacturers looking at major investments into capacity. The perceived bottleneck in
downstream processing can be addressed with lower-risk approaches, such as streamlining current production processes, with
moderate-risk approaches, such as introducing technologies that have already proven suitable in other industry settings, or
with higher-risk approaches involving the incorporation of novel technologies.
In several cases, these novel technologies have already proven their credentials in several processes and companies following
the paths set by the first adopters, the trailblazers of the industry, can be assured that the technologies involved now have
established their credibility.
The future of biomanufacturing is likely to rely more on innovation and flexibility than on traditional strengths, such as
large facilities and the financial muscle to invest in them. Disposable manufacturing is likely to play an increasingly important
role as companies maneuver in a crowded market to protect their R&D investments while more and more generics become available.
The ability to scale up or down quickly, to switch to new campaigns rapidly and to produce multiple products in the same facility
will be a key metric of success. Ultimately, the future of bioprocessing will require industry players to embrace the need
to change.
Uwe Gottschalk, PhD, is vice-president of purification technologies at Sartorius Stedim Biotech GmbH, uwe.gottschalk@sartorius.com
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