Chromatography, ion exchange

Cramer, Steven M.; Natarajan, Venkatesh

doi:10.1002/0471250589.ebt051

Cited by 2 publications

(5 citation statements)

References 75 publications

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“…A charged stationary phase will interact, bind, and separate an oppositely charged macromolecule such as a protein or DNA present in the mobile phase. Cation exchange occurs where the stationary phase is negatively charged and the macromolecule in the mobile phase is positively charged 1, 2. An important parameter to assess column performance, when considering this type of chromatographic operation is breakthrough or frontal chromatography, i.e., the process where the column becomes saturated with solute.…”

Section: Introductionmentioning

confidence: 99%

“…An important parameter to assess column performance, when considering this type of chromatographic operation is breakthrough or frontal chromatography, i.e., the process where the column becomes saturated with solute. Large‐scale columns are usually run to maximize yield by reduction of losses during loading as determined from the breakthrough curve 2…”

Section: Introductionmentioning

confidence: 99%

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Design and characterization of a microfluidic packed bed system for protein breakthrough and dynamic binding capacity determination

Shapiro

Haswell

Lye

et al. 2009

Biotechnology Progress

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A 1.5 microL ion exchange chromatography column to accommodate resins used for biopharmaceutical processing has been designed to produce breakthrough curves and to quantify dynamic and maximum protein binding capacities. Channels within a glass chip were fabricated using photolithography and isotropic etching. The design includes a 1 cm long microfluidic column in which compressible, polydispersed porous agarose beads (70 mum mean diameter) were packed using a keystone method where particles aggregate in a narrow channel. The depth of the column is such that two bead layers exist. The fabrication technique used forms Cartesian geometries as opposed to circular cross sections found in standard columns. The voidage was therefore higher than standard values when measured by 3D confocal microscopy. In conjunction with microscopic techniques, the column allows visualization of events within the bed such as adsorption profiles that would otherwise be difficult to observe. In this work, the binding of fluorescently labeled protein during isocratic loading was used to generate breakthrough from the microcolumn. Useful breakthrough curves were achieved using mobile phase velocities from 60 to 270 cm h(-1). Calculated dynamic binding capacities were compared well with previously published data on conventional scale columns. The microfluidic chromatography column described here thus allows study of process scale chromatography behavior at scales 20,000 times smaller than in current practice. The work described in this article is representative of the proof of principle of a potentially powerful tool for the generation of microfluidic process bed data for the biopharmaceutical industry.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Design and characterization of a microfluidic packed bed system for protein breakthrough and dynamic binding capacity determination

Shapiro

Haswell

Lye

et al. 2009

Biotechnology Progress

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“…We will have much to say about this in our discussion of design strategy. As this review is being written, fixed-bed sorption operations, in particular various forms of chromatography (Adams et al, 1999;Cramer and Natarajan, 1999;Guiochon et al, 1994;Kaarsnaes, 1999;Lightfoot, 1999;McGuire and Bothwell, 1999;Rathore and Velayudhan, 2003;Roth and Yarmush, 1999;Thoemmes, 1999) are dominant for recovery of valuable products from impure dilute solutions and for separations of closely related materials. They will be discussed in some detail below.…”

Section: Final Purificationmentioning

confidence: 99%

“…A great variety of sorbents is now available (Adams et al, 1999), and chief among them for preparative purposes are size exclusion (SEC; Roth and Yarmush, 1999), ionexchange (IX; Cramer and Natarajan, 1999), hydrophobic interaction (HIC; Kaarsnaes, 1999), and ''affinity'' (Stahl et al, 1999) adsorbents. There are a great many subtypes within each group, including for example, both weak and strong acid and base exchangers -and many gradations of affinity systems.…”

Section: Practical Considerationsmentioning

confidence: 99%

“…It is perhaps the most widely used operating mode for protein fractionation. A common example would be the addition of increasing amounts of salt in the feed to an ion-exchange column, but this general process can be used for any of the adsorbent types previously described (Cramer and Natarajan, 1999;Kaarsnaeas, 1999;Stahl et al, 1999). This process depends on a species-dependent response of the adsorption equilibrium to elutant concentration, and it thus adds a new separation mechanism to the selective migration discussed previously: selective elution (Coffman et al, 1994;Yamamoto et al, 1993;Yamamoto, 1995).…”

Section: Gradient Elutionmentioning

confidence: 99%

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Bioseparations

Lightfoot

Moscariello

2004

Biotech & Bioengineering

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Here we review key applications of separation technology in applied biology. We first sketch out the field as a whole, but then narrow our scope to the processing of fermentation products, particularly to high-value biologicals such as proteins and nucleotides. We go on to provide a qualitative overview describing the importance and general nature of this large field, major trends, and the strategies that have proven most fruitful in evolving effective separation and purification processes. We then give a detailed description of individual separations equipment and the principles governing their operation. We concentrate throughout on making the available literature accessible to the reader; we provide what is hoped to be a representative set of basic references. However, these references, in turn, include some that suggest promising new developments as well as a number of more specialized reviews. We hope that our overall result provides the reader with access to the most relevant literature. B

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Chromatography, ion exchange

Cited by 2 publications

References 75 publications

Design and characterization of a microfluidic packed bed system for protein breakthrough and dynamic binding capacity determination

Design and characterization of a microfluidic packed bed system for protein breakthrough and dynamic binding capacity determination

Bioseparations

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