Prediction of the performance of preparative affinity chromatography

Chase, Howard A.

doi:10.1016/s0021-9673(01)89041-5

Cited by 411 publications

(231 citation statements)

References 13 publications

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“…A similar tendency of 450 dynamic adsorption capacity was obtained in accordance with the results observed by 451 static adsorption experiments. Dynamic processes result mostly in a worse 452 performance than a static one; the degree strongly depends on the operation 453 conditions [4,40]. The dynamic adsorption capacity of GOD was also reduced after Table 3 studies show that TC-mPEG2k exhibits much better adsorption performance than 459 TC-mPEG5k.…”

Section: Affinity Chromatography Of Glucose Oxidase Onto Concanavalinmentioning

confidence: 99%

Preparation and characterization of PEGyated Concanavalin A for affinity chromatography with improved stability

Wen

Niemeyer

2011

Journal of Chromatography B

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22In order to improve its stability, immobilized Concanavalin A (Con A) on Toyopearl 23 adsorbents was conjugated with Monomethoxy poly(ethylene glycol) succinimidyl 24propionate (mPEG-SPA) with different molecular weight. Colorimetric method using 25 ninhydrin to determine the PEGylated degree was first proposed in this study, which 26 has proved to be easy applicable and reproducible. The PEGylation reaction was 27 studied in detail to elucidate how the parameters such as reaction time, pH value, 28 molar ratio of mPEG-SPA to Con A, and molecular weight of mPEG-SPA affect the 29 PEGylated degree. The adsorption isotherms of glucose oxidase (GOD) onto native 30 and PEGylated Con A adsorbents showed that the modification did not alter 31 substantially the specificity of the carbohydrate binding ability of Con A. However, 32 the binding capacity for GOD was slightly reduced probably due to the steric 33 hindrance caused by mPEG chains. The adsorption kinetic studies revealed the lower 34 adsorption rate after PEGylation which was still attributed to the steric effect.

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Section: Affinity Chromatography Of Glucose Oxidase Onto Concanavalinmentioning

confidence: 99%

Preparation and characterization of PEGyated Concanavalin A for affinity chromatography with improved stability

Wen

Niemeyer

2011

Journal of Chromatography B

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“…Equation (4) is the Langmuir isotherm which is frequently used for describing the adsorption behaviour of proteins to a variety of adsorbents, including ion exchange and affinity chromatography media. A kinetic rate constant model has been introduced to describe the adsorption of proteins to affinity chromatography media by one single adsorption rate constant [21]. In this approach, the rate of mass transfer of the target molecule to the stationary phase is described by Eq.…”

Section: Lumped Kinetic Modelmentioning

confidence: 99%

Adsorption of plasmid DNA on anion exchange chromatography media

Tarmann

Jungbauer²

2008

J of Separation Science

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Original PaperAdsorption of plasmid DNA on anion exchange chromatography media Anion exchange chromatography (AEC) is a useful and effective tool for DNA purification, but due to average pore sizes between 40 and 100 nm most AEC resins lack truly useful binding capacities for plasmid DNA (pDNA). Equilibrium binding capacities and uptake kinetics of AEC media including conventional media (Source 30 Q, Q Sepharose HP), a polymer grafted medium (Fractogel EMD DEAE (M)), media with large pores (Celbeads DEAE, PL SAX 4000 30 lm) and a monolithic medium (CIM-DEAE) were investigated by batch uptake or shallow bed experiments at two salt concentrations. Theoretical and experimental binding capacities suggest that the shape of the pDNA molecule can be described by a rod with a length to diameter ratio of 20:1 and that the molecule binds in upright position. The arrangement of DNA like a brush at the surface can be considered as entropy driven, kind of selfassembly process which is inherent to highly and uniformly charged DNA molecules. The initial phase of adsorption is very fast and levels off, associated with a change in mass transfer mechanism. Feed concentrations higher than 0.1 mg/mL pDNA pronounce this effect. Monolithic media showed the fastest adsorption rate and highest binding capacity with 13 mg pDNA per mL.

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“…The sensitivity of the chromatography step yield to ligand density was relatively strong, as a 3-fold change in peptide density was sufficient to increase step yields from less than 10% to greater than 90% (Kelley et al, 2004). This required a careful titration of ligand density, the results of which were supported by the use of a mathematical model of the chromatographic separation using Simulus software (Chase, 1984;Wiblin et al, 1995); the simulation provides a reasonable fit to the performance of the chromatography step based on input data including the peptide-protein dissociation constant, binding kinetics, peptide density, flowrate, and product concentration in the load (data not shown).…”

Section: Optimization Of Tn82 Peptide Densitymentioning

confidence: 99%

Development and validation of an affinity chromatography step using a peptide ligand for cGMP production of factor VIII

Kelley¹,

Tannatt²,

Magnusson

et al. 2004

Biotech & Bioengineering

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An affinity chromatography step was developed for purification of recombinant B-Domain Deleted Factor VIII (BDDrFVIII) using a peptide ligand selected from a phage display library. The peptide library had variegated residues, contained both within a disulfide bond-constrained ring and flanking the ring. The peptide ligand binds to BDDrFVIII with a dissociation constant of f1 AM both in free solution and when immobilized on a chromatographic resin. The peptide is chemically synthesized and the affinity resin is produced by coupling the peptide to an agarose matrix preactivated with N-hydroxysuccinimide. Coupling conditions were optimized to give consistent and complete ligand incorporation and validated with a robustness study that tested various combinations of processing limits. The peptide affinity chromatographic operation employs conditions very similar to an immunoaffinity chromatography step currently in use for BDDrFVIII manufacture. The process step provides excellent recovery of BDDrFVIII from a complex feed stream and reduces host cell protein and DNA by 3 -4 logs. Process validation studies established resin reuse over 26 cycles without changes in product recovery or purity. A robustness study using a factorial design was performed and showed that the step was insensitive to small changes in process conditions that represent normal variation in commercial manufacturing. A scaled-down model of the process step was qualified and used for virus removal studies. A validation package addressing the safety of the leached peptide included leaching rate measurements under process conditions, testing of peptide levels in product pools, demonstration of robust removal downstream by spiking studies, end product testing, and toxicological profiling of the ligand. The peptide ligand affinity step was scaled up for cGMP production of BDDrFVIII for clinical trials. B 2004 Wiley Periodicals, Inc.

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Prediction of the performance of preparative affinity chromatography

Cited by 411 publications

References 13 publications

Preparation and characterization of PEGyated Concanavalin A for affinity chromatography with improved stability

Preparation and characterization of PEGyated Concanavalin A for affinity chromatography with improved stability

Adsorption of plasmid DNA on anion exchange chromatography media

Development and validation of an affinity chromatography step using a peptide ligand for cGMP production of factor VIII

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