Continuous separation of green fluorescent protein by annular chromatography

Uretschläger, Andrea; Einhauer, A.; Jungbauer, Alois

doi:10.1016/s0021-9673(00)00833-5

Cited by 19 publications

(6 citation statements)

References 8 publications

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“…Therefore, the purification of pharmaceutical biocompounds is usually complex and based in multi-step platforms [22]. For example, the current strategies for GFP purification involve, mainly, the use of multiple chromatographic units, which are complex and costly [18,[23][24][25][26][27].…”

Section: Introductionmentioning

confidence: 99%

Economic analysis of the production and recovery of green fluorescent protein using ATPS-based bioprocesses

Torres‐Acosta

Santos

Ventura

et al. 2021

Separation and Purification Technology

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Section: Introductionmentioning

confidence: 99%

Economic analysis of the production and recovery of green fluorescent protein using ATPS-based bioprocesses

Torres‐Acosta

Santos

Ventura

et al. 2021

Separation and Purification Technology

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“…More recently, continuous annular chromatography (CAC) became commercially available (P-CAC, preparative continuous annular chromatography; Prior Separation Technology, Austria). Since then it has been applied for protein separation by size-exclusion Iberer et al, 2001;Uretschlaeger et al, 2001), as well as experimental capture of an antibody by ion exchange (Giovannini and Freitag, 2001). …”

Section: Isolation Of Protein Therapeuticsmentioning

confidence: 99%

Continuous annular chromatography: General characterization and application for the isolation of recombinant protein drugs

Vogel

Nguyen

Pritschet³

et al. 2002

Biotech & Bioengineering

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Isolation of recombinant protein drugs from cell culture supernatant is usually performed in batch mode, even if the fermentation process itself is continuous. As a novel approach, continuous separation techniques like continuous annular chromatography (CAC) can be used for continuous isolation. The potential of CAC for industrial application is demonstrated by continuous isolation of rFVIII from cell culture supernatant in pilot scale (i.e., 144-288 l/day). Thirty-fold concentration can be achieved at 94% yield, while purity is increased 3-5-fold. For this a batch direct feed ion exchange chromatography method was adapted to a commercial preparative CAC system (P-CAC). A headspace loading technique was used to maximize the concentration factor, while buffer incompatibility problems were addressed by a specifically modified inlet geometry. To allow sterile on-line coupling to FVIII-producing perfusion fermenters, an autoclavable pilot scale P-CAC prototype was developed. General characterization of P-CAC revealed a current limitation of the technology, i.e., variations in the outlet flow rates of up to +/-20%. These flow variations are shown to be caused mainly by a nonuniform annular resin bed and in turn result in "peak wobbling," i.e., the slight variation of peak position (up to +/-4 degrees ) and shape (e.g., A(s) = 0.9-1.4) as a specific function of column position. Some additional peak broadening, although less significant, is caused by a "peak oscillation" effect that results from the necessary segmentation of flow into discrete outlets. Both effects are only measurable if peaks are either monitored continuously or at least measured at multiple column positions. For isolation processes, these nonideal flow phenomena mean that more outlet streams have to be collected in order to achieve maximum yield and thus the achievable concentration factor is somewhat lower than the theoretical maximum.

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“…11a-f. Components of a P-CAC system: a Teflon ring; b metal plate with 90 exit holes, combining inner and outer cylinder; c metal plate with filter and filter clamp on Teflon ring; d inner cylinder made of PVC; e outer cylinder made of glass; f entire P-CAC system fluorescent protein [42] and IgG [26]. Green fluorescent protein was used as a model substance in a separation experiment where two separation resins were packed into one column of a CAC [43].…”

Section: Applicationsmentioning

confidence: 99%

Preparative continuous annular chromatography (P-CAC), a review

Uretschläger

Jungbauer

2002

Bioprocess and Biosystems Engineering

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Notation c liquid phase solute concentration (g/l) c* liquid concentration in equilibrium (g/l) D z axial dispersion coefficient (cm 2 /min) D m solute dispersion coefficient in the axial direction (cm 2 /min) D h solute dispersion coefficient in the angular direction (cm 2 /min) k 0 a global mass transfer parameter (s -1 ) k¢ A normalized retention of compound A k¢ B normalized retention of compound B K distribution coefficient K m mass transfer coefficient between the mobile and the stationary phases K eq equilibrium partition coefficient m p amount of pure protein (mg) n cycle factor N eff effective plate number N th number of theoretical plates P productivity (mg/(ml min)) q solid phase solute concentration (g/l) q* solid concentration in equilibrium (g/l) Q F feed flow rate (ml/min) Q E eluent flow rate (ml/min) Q T total flow rate (ml/min) Q quantity Q R recovery ratio R resolution S particle area per unit column volume (cm 2 /ml) t retention time (min) t max reduced retention time (min) t t max retention time of dextran blue (min) t F feed time (min) t c cycle time (min) t c,1 time for the first cycle of batch column (min) t c,b time for n cycles of batch column (min) t¢ c,1 time for the first cycle of P-CAC (min) t¢ c,c time for n cycles of P-CAC (min) u superficial velocity (cm/min) V B buffer consumption V t column volume (ml) V E elution volume (ml)

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Continuous separation of green fluorescent protein by annular chromatography

Cited by 19 publications

References 8 publications

Economic analysis of the production and recovery of green fluorescent protein using ATPS-based bioprocesses

Economic analysis of the production and recovery of green fluorescent protein using ATPS-based bioprocesses

Continuous annular chromatography: General characterization and application for the isolation of recombinant protein drugs

Preparative continuous annular chromatography (P-CAC), a review

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