and in particular in the development of chromatographic separations, the techniques employed are separated into batch and packed
bed column chromatography [8, 9]. Recently, applications have
been developed that employ both diffusive and convective media,
but the latter are still missing a commercially available flow mode
based HT technology. While batch HT methods allow the evaluation of multiple conditions in parallel, including binding capacity
measurements from small product masses due to the low volumes
of stationary phases employed. Moreover, the miniature packed
bed column technique (i.e., RoboColumns) can be easily scaled
up or down [10]. It can also return an efficient way of evaluating
different stationary phases due to the advantage of packing any
beaded based resin at a cost. This is an application that is traditionally regarded to be better accomplished with batch methods,
requiring, however, sophisticated processes for preparing resin
slurry multiwell plates [11]. Jacob and coworkers [12] have
demonstrated that it is possible to design and synthesize affinity
resins for high recovery of novel influenza antigens and various
recombinant proteins (erythropoietin, immunoglobulins, etc.)
[13, 14]. Therefore, such HT techniques can be employed to
evaluate their performance.
In this instance, the scale-down performance of the miniature
columns was leveraged to guide development efforts in a multivariate input space, including stationary phase type and operating
conditions, for recombinant vaccine antigen purification. The
approach relies heavily on the performance of the miniature columns and as such it employs custom solutions and in-house developed tools that deliver full walk-away automation across the
different parts of the HT study, that is, buffer preparation, method
definition, experiment completion, and results reporting. The followed workflow employs a Tecan Freedom EVO
® 200 automated
liquid handling station (Tecan Group Ltd., M€ annedorf, Switzerland) and MATLAB
® (The MathWorks, MA, USA) codes, compiled into executables, which convert user-defined inputs into
robotic commands. These are then executed within generic Tecan
Freedom EVOware
® (Tecan Group Ltd.) based scripts that implement all necessary actions to complete a study with no end-user
intervention [10]. Here, both 0.2 and 0.6 mL columns, packed
with various types of resin, can be used in flowthrough or bind and
elute mode, with the latter employing isocratic, multistep, and
multislope gradient elution. The workflow is demonstrated by
implementing an early purification strategy in scouting for multimodal resins and conditions for two different strains of rotavirus
antigen. A small number of well-planned experiments were sufficient to elucidate a lead resin candidate and separation conditions
which were then found to be scalable to a larger pilot scale. Hence,
the combination of HT techniques, automation, and a systematic
screening approach can lead to attractive and feasible purification
process conditions in a rapid fashion.
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Shaleem I. Jacob et al.
bed column chromatography [8, 9]. Recently, applications have
been developed that employ both diffusive and convective media,
but the latter are still missing a commercially available flow mode
based HT technology. While batch HT methods allow the evaluation of multiple conditions in parallel, including binding capacity
measurements from small product masses due to the low volumes
of stationary phases employed. Moreover, the miniature packed
bed column technique (i.e., RoboColumns) can be easily scaled
up or down [10]. It can also return an efficient way of evaluating
different stationary phases due to the advantage of packing any
beaded based resin at a cost. This is an application that is traditionally regarded to be better accomplished with batch methods,
requiring, however, sophisticated processes for preparing resin
slurry multiwell plates [11]. Jacob and coworkers [12] have
demonstrated that it is possible to design and synthesize affinity
resins for high recovery of novel influenza antigens and various
recombinant proteins (erythropoietin, immunoglobulins, etc.)
[13, 14]. Therefore, such HT techniques can be employed to
evaluate their performance.
In this instance, the scale-down performance of the miniature
columns was leveraged to guide development efforts in a multivariate input space, including stationary phase type and operating
conditions, for recombinant vaccine antigen purification. The
approach relies heavily on the performance of the miniature columns and as such it employs custom solutions and in-house developed tools that deliver full walk-away automation across the
different parts of the HT study, that is, buffer preparation, method
definition, experiment completion, and results reporting. The followed workflow employs a Tecan Freedom EVO
® 200 automated
liquid handling station (Tecan Group Ltd., M€ annedorf, Switzerland) and MATLAB
® (The MathWorks, MA, USA) codes, compiled into executables, which convert user-defined inputs into
robotic commands. These are then executed within generic Tecan
Freedom EVOware
® (Tecan Group Ltd.) based scripts that implement all necessary actions to complete a study with no end-user
intervention [10]. Here, both 0.2 and 0.6 mL columns, packed
with various types of resin, can be used in flowthrough or bind and
elute mode, with the latter employing isocratic, multistep, and
multislope gradient elution. The workflow is demonstrated by
implementing an early purification strategy in scouting for multimodal resins and conditions for two different strains of rotavirus
antigen. A small number of well-planned experiments were sufficient to elucidate a lead resin candidate and separation conditions
which were then found to be scalable to a larger pilot scale. Hence,
the combination of HT techniques, automation, and a systematic
screening approach can lead to attractive and feasible purification
process conditions in a rapid fashion.
120
Shaleem I. Jacob et al.