The influence of cell adsorbent interactions on protein adsorption in expanded beds

Fernández-Lahore, Hector Marcelo; Geilenkirchen, S.; Boldt, Klaus; Nagel, Alexey; Kula, Maria‐Regina; Thömmes, Jörg

doi:10.1016/s0021-9673(99)01308-4

Cited by 78 publications

(46 citation statements)

References 11 publications

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“…2, a maximum surface coverage (percentage of maximum capacity) of 39% is achieved, meaning that 61% of total surface sites are unoccupied at this stage. The equilibrium capacity obtained is of the same order of magnitude as the reported value of approximately 500 × 10 6 cells/mL of gel for the adsorption of S. cereVisiae to a copper-primed Streamline Chelating adsorbent in finite bath mode (23).…”

Section: Resultssupporting

confidence: 75%

“…The kinetics model proposed by Daniels (27) used in other cell adsorption studies (23,24) did not represent well our experimental data (results not shown). As verified by Lin et al (24), it is believed that the Daniels model better describes systems showing strong biomass-adsorbent interactions.…”

Section: Resultscontrasting

confidence: 58%

“…Various methods for the evaluation of biomass-adsorbent interaction have been reviewed (22). Cell adsorption experiments can be performed by the finite bath method (23,24) and by expanded bed operation (25,26) in order to take into account all hydrodynamic and physicochemical aspects.…”

Section: Introductionmentioning

confidence: 99%

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A Study of the Interaction of HEK-293 Cells with Streamline Chelating Adsorbent in Expanded Bed Operation

et al. 2008

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Expanded bed adsorption (EBA) is an efficient protein purification process reducing time and steps of downstream processing (DSP) since nonclarified culture media can be processed directly without prior treatments such as filtration or centrifugation. However, cells and debris can interact with the adsorbent and affect bed stability as well as purification performance. To optimize EBA operating conditions these biomass/adsorbent interactions have to be understood and characterized. The adsorption of Human Embryonic Kidney cells (HEK 293) on unprimed and nickel-primed metal affinity adsorbent was studied in a closed loop EBA setup. With the unprimed adsorbent, the overall level of interaction observed was nonsignificant. With the nickel-primed adsorbent and an initial cell concentration ranging from 0.08 × 10 6 to 0.2 × 10 6 cells/mL, biomass/adsorbent interaction was found to be moderate and the adsorption apparent first-order kinetic rate constant was determined to be k ) 0.009 to 0.011 min -1 .

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Section: Resultssupporting

confidence: 75%

Section: Resultscontrasting

confidence: 58%

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A Study of the Interaction of HEK-293 Cells with Streamline Chelating Adsorbent in Expanded Bed Operation

et al. 2008

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“…Second, interaction of the biomass with the adsorbent base matrix or ligands may interfere with product adsorption and expanded bed hydrodynamic stability. This detrimental phenomenon occurs for a variety of cell/resin combinations (Fernandez-Lahore et al, 2000;Feuser et al, 1999a;1999b;Lin et al, 2001). Most notably, bacterial and yeast cells complex with anion exchange resins, while hybridoma cells do so with cation exchangers.…”

Section: Introductionmentioning

confidence: 99%

Compatibility of column inlet and adsorbent designs for processing of corn endosperm extract by expanded bed adsorption

Menkhaus

Glatz²

2004

Biotech & Bioengineering

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Corn has emerged as a viable host for expression of recombinant proteins; targeted expression to the endosperm has received particular attention. The protein extracts from corn endosperm differ from those of traditional hosts in regard to the nature of residual solids and extracted matrix contaminants. Each of these differences presents reasons for considering expanded bed adsorption for product capture and new considerations for limitations of the method. In this work three inlet-flow distribution devices (mesh, glass ballotini, and localized mixing) and six adsorbents with different physical (size and density), chemical (ligand), and base matrix properties were evaluated to determine conditions compatible with processing of crude corn endosperm extract by expanded bed adsorption.Of the inlet devices evaluated, the design with localized mixing at the inlet (as produced commercially by UpFront Chromatography A/S, Copenhagen, DK) allowed solids up to 550 Am into the column without clogging for all flow rates evaluated. A mesh at the inlet with size restriction of either 50 Am or 80 Am became clogged with very small corn particles (< 44 Am). When glass ballotini was used, large particles (550 Am) passed through for high flow rates (570 cm/h), but even small (< 44 Am) particles became trapped at a lower flow rate (180 cm/h). The physical and chemical properties of the resin determined whether solids could be eluted. The denser UpFront adsorbents allowed for complete elution of larger and more concentrated corn solids than the currently available Amersham Streamline adsorbents (Amersham Biosciences, Piscataway, NJ) as a result of the former's higher flow rate for the desired 2Â expansion (570 cm/h for UpFront vs. 180 cm/h for Streamline). All corn solids < 162 Am eluted through nonderivatized UpFront resin. Larger corn solids began to accumulate due to their elevated sedimentation velocities. Feeds of < 44 Am solids at 0.45% and 2.0% dry weight successfully eluted through ion exchange adsorbents (DEAE and SP) from UpFront. However, significant accumulation occurred when the solids size increased to a feed of < 96 Am solids, thus indicating a weak interaction between corn solids and both forms of ion exchange ligands. Expanded beds operated with Streamline ion exchange adsorbents (DEAE and SP) did not allow full elution of corn solids of < 44 Am. A hyperdiffuse style EBA resin produced by Biosepra (Ciphergen Biosystems, Fremont, CA) with CM functionality showed a severe interaction with corn solids that collapsed the expanded bed and could not be eliminated with elevated flow rates or higher salt concentration. B 2004 Wiley Periodicals, Inc.

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“…Recently, a set of three methods (finite bath adsorption, biomass pulse response experiment, and residence-time distribution analysis) has been established in our laboratories to identify, evaluate, and minimize the biomass/ adsorbent interactions in a logical and strategic way (Fernández-Lahore et al, 1999, 2000Feuser et al, 1999;Lin et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

The influence of biomass on the hydrodynamic behavior and stability of expanded beds

Lin

Thömmes

Kula

et al. 2004

Biotech & Bioengineering

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Expanded bed adsorption is an innovative chromatographic technology that allows the introduction of particle-containing feedstock without the risk of blocking the bed. Provided a perfectly classified fluidized bed (termed expanded bed) is formed in the crude feedstock and the biomass is not influencing protein transport towards the adsorbent surface, a sorption performance comparable to packed beds is found. The influence of biomass on the hydrodynamic stability of expanded beds is essential and was investigated systematically in this article. Residencetime distribution analyses were performed using model systems and a yeast suspension under various fluid-phase conditions. It is demonstrated that three factors (biomass/ adsorbent interactions, biomass concentration, and flow rate) play an interdependent role disturbing the classified fluidization of an expanded bed. A clear correlation between the degree of aggregative fluidization-obtained by PDE modeling of RTD data-and the expansion behavior of the fluidized bed has been found. Thus, combining three analytical methods, namely cell transmission index analysis, expansion analysis, and RTD analysis provides a solid base for understanding and control of the fluidization behavior and thus further process design during the initial phase of process development. B 2004 Wiley Periodicals, Inc.

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The influence of cell adsorbent interactions on protein adsorption in expanded beds

Cited by 78 publications

References 11 publications

A Study of the Interaction of HEK-293 Cells with Streamline Chelating Adsorbent in Expanded Bed Operation

A Study of the Interaction of HEK-293 Cells with Streamline Chelating Adsorbent in Expanded Bed Operation

Compatibility of column inlet and adsorbent designs for processing of corn endosperm extract by expanded bed adsorption

The influence of biomass on the hydrodynamic behavior and stability of expanded beds

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