Purification of proteins by adsorption chromatography in expanded beds

Chase, Howard A.

doi:10.1016/0167-7799(94)90046-9

Cited by 323 publications

(148 citation statements)

References 16 publications

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“…Expanded Bed Adsorption (EBA) is a chromatographic technique for separation and purification of biological products directly from crude feedstock, without centrifugation, microfiltration and other prior clarification steps (Chase, 1994;Hjorth;Thömmes, 1997;Anspach et al, 1999;Trinh et al, 2006;Camprubi et al, 2006;Tan et al, 2006). This technique permits crude feeding into the chromatographic column without initial treatment and, as the bed expands, it increases adsorbent surface contact, making interaction with the target molecule more effective (Roy et al, 1999;Fernandez-Lahore et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Expanded bed adsorption of bromelain (E.C. 3.4.22.33) from Ananas comosus crude extract

Silveira

Souza-Jr²,

Santana

et al. 2009

Braz. J. Chem. Eng.

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-This work focuses on the adsorption of Bromelain in expanded bed conditions, such as the adsorption kinetics parameters. The adsorption kinetics parameters showed that after 40 minutes equilibrium was achieved and maximum adsorption capacity was 6.11 U per resin mL. However, the maximum adsorption capacity was only determined by measuring the adsorption isotherm. Only by the Langmuir model the maximum adsorption capacity, Qm, and dissociation constant, kd, values could be estimated as 9.18 U/mL and 0.591, respectively, at 25 °C and 0.1 mol/L phosphate buffer pH 7.5. A column made of glass with an inner diameter of 1 cm was used for the expanded bed adsorption (EBA). The residence time was reduced 10 fold by increasing the expansion degree 2.5 times; nonetheless, the plate number (N) value was reduced only 2 fold. After adsorption, the bromelain was eluted in packed bed mode, with a downward flow. The purification factor was about 13 fold and the total protein was reduced 4 fold. EBA showed to be feasible for purification of bromelain.

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Section: Introductionmentioning

confidence: 99%

Expanded bed adsorption of bromelain (E.C. 3.4.22.33) from Ananas comosus crude extract

Silveira

Souza-Jr²,

Santana

et al. 2009

Braz. J. Chem. Eng.

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“…It combines clarification (replacing centrifugation or microfiltration), concentration (replacing ultrafiltration), and chromatographic separation (replacing packed bed chromatography) into a single unit operation. The customized size and/or density distribution of the EBA resins allow for stable fluidization of the chromatographic bed (minimal back mixing of both stationary and mobile phases) with bed expansion that allows passage of feed solids (Chase, 1994;Hjorth, 1997;Lihme et al, 2000). This technique has been successful with a variety of feeds, including culture broths, homogenates, milk, and some plant extracts (Bai and Glatz, 2003b and references therein).…”

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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“…It is of importance to elucidate the expanded bed hydrodynamics for a better EBA process design. 5,6,18 Thus, the axial dispersion behavior in the expanded bed of the cation resin was investigated to confirm whether it is suitable for EBA operation.…”

Section: Axial Liquid-phase Dispersion Behavior In the Expanded Bedmentioning

confidence: 99%

“…As an innovative chromatography mode, expanded bed adsorption (EBA) has great advantages over conventional packedbed chromatography. 5,6 EBA is a single-pass unit operation in which products are recovered and concentrated from crude feedstock and transferred to a process for further chromatographic purification, without the need for feedstock clarification. Over the past decade, a number of successful applications using EBA with a variety of biological systems, such as whole yeast, mammalian cells and their homogenates and so on, have been reported in the literature.…”

Section: Introductionmentioning

confidence: 99%

Hydrodynamics in an expanded bed of large size ion‐exchange resin, and natural product adsorption

Chen

Wang

et al. 2007

J of Chemical Tech & Biotech

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Successful applications of expanded bed adsorption (EBA) technology have been widely reported in the literature for protein purification. Little has been reported on the recovery of natural products and active components of Chinese herbal preparations using EBA technology. In this study, the hydrodynamic behavior in an expanded bed of cation resin, 001 × 7 Styrene-DVB, was investigated. Ephedrine hydrochloride (EH) was used as a model natural product to test the dynamic binding capacity (DBC) in the expanded bed. EBA of EH directly from a feedstock containing powdered herbs has also been investigated. These particles are different from commercially available expanded bed adsorbents by virtue of their large size (205 to 1030 µm). When the adsorbent bed is expanded to approximately 1.3 to 1.5 times its settled bed height, the axial liquid-phase dispersion coefficient was found to be of the order 10 −5 m 2 s −1 , which falls into the range 1.0 × 10 −6 to 1.0 × 10 −5 m 2 s −1 observed previously in protein purification. Because of the favorable column efficiency (low axial dispersion coefficient), the recovery yield and purification factor values of EH directly from a feedstock reached 86.5% and 18, respectively. The results suggest that EBA technology holds promise for the recovery of natural products and active components of Chinese herbal preparations.

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Purification of proteins by adsorption chromatography in expanded beds

Cited by 323 publications

References 16 publications

Expanded bed adsorption of bromelain (E.C. 3.4.22.33) from Ananas comosus crude extract

Expanded bed adsorption of bromelain (E.C. 3.4.22.33) from Ananas comosus crude extract

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

Hydrodynamics in an expanded bed of large size ion‐exchange resin, and natural product adsorption

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