Short fibrous supports for preparative chromatographic separations of biomolecules

King, Jy-Kung; Pinto, Neville G.

doi:10.1016/0021-9673(92)80149-o

Cited by 22 publications

(10 citation statements)

References 20 publications

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“…Mechanical stability, minimal porosity, and the ability to operate at relatively low back pressures were realized, with static BSA binding capacities of 115 mg g −1 of fabric, which reduced by a factor of 6 at linear velocities up to ∼54 cm min −1 . Pinto and coworkers extended the works of Wikstrom and Larson, using polysulfone (PS) as the base polymer in randomly packed, short‐fiber columns for anion exchange separation of BSA and β‐lactoglobulin . The results of the latter study showed that equilibrium protein adsorption capacities of PS fibers depended on the size of protein, pH, and salt concentration; maximizing at ∼20 and 400 mg g −1 fiber, respectively.…”

Section: Introductionmentioning

confidence: 97%

“…In addition to the more common support/stationary phases described above, there have been attempts to develop fiberbased chromatographic systems for bioseparations. [9][10][11][12][13][14][15][16][17][18][19] As described by Marcus,20,21 there are practical reasons to investigate fibrous phases for protein separations, these include low-cost readily available materials, the ability to affect multiple column formats, a diversity of native and functionalized surfaces, and excellent fluidics and mass transfer characteristics. Natural and synthetic polymer fibers have found use in a variety of chromatographic modes.…”

Section: Introductionmentioning

confidence: 99%

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Initial evaluation of protein throughput and yield characteristics on nylon 6 capillary‐channeled polymer (C‐CP) fiber stationary phases by frontal analysis

Randunu

Marcus

2013

Biotechnology Progress

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Nylon 6 capillary-channeled polymer (C-CP) fibers are investigated as an alternative support/stationary phase for downstream processing of macromolecules. Ionizable amine and carboxylic acid end groups on the native fiber surface allow for ion exchange chromatography (IEC). The low cost and ability to operate at high linear velocities and low back pressures are practical advantages of C-CP fibers for preparative-scale macromolecule separations. The lack of fiber porosity ensures facile adsorption/desorption that is conducive to high throughput and recoveries/yields. Described here is a preliminary investigation of the processing characteristics of lysozyme on nylon 6 fibers with an eye toward downstream processing applications. Fibers were packed into microbore (0.8 mm i.d.) and analytical-size (2.1 mm i.d.) columns for the evaluation of the role of linear velocity on pressure drop, frontal throughput, and yield. Protein isolation by frontal development involved three steps: loading of the column to breakthrough, an aqueous wash, and a salt wash to recover the protein. Frontal throughput was evaluated with different salt concentrations (0-1000 mM NaCl) and different linear velocities (6-24 mm s(-1)). The observed throughput values are in the range of 0.12-0.20 mg min(-1) when 0.25 mg mL(-1) lysozyme (in 20 mM Tris-HCl) is loaded onto 78 mg of C-CP fiber in 0.52 mL volume analytical columns. Increased throughput and yield were found when protein was loaded and eluted at high linear velocity. Results of this study lend credence to the further development of C-CP fibers for biomacromolecule processing on larger scales.

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Section: Introductionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Initial evaluation of protein throughput and yield characteristics on nylon 6 capillary‐channeled polymer (C‐CP) fiber stationary phases by frontal analysis

Randunu

Marcus

2013

Biotechnology Progress

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“…This work was actually an extension of previous works on silica RPSF supports by that group and others [16,17]. The specific application here was the anionexchange separation of the proteins BSA and b-lactoglobulin.…”

Section: Separations On Fiber Staple Phasesmentioning

confidence: 92%

Use of polymer fiber stationary phases for liquid chromatography separations: Part II – applications

Marcus¹

2009

J of Separation Science

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The idea of using polymer fibers as stationary phases for LC is not a new one. There are in fact a number of good reasons for which they should be considered. Presented here are a number of applications of polymer fiber stationary phases for chemical separations. Fibers placed in columns are employed in three basic formats: staple, whole fabrics, and aligned fibers. Natural and synthetic fibers have found use in a variety of chromatographic modes including RP, size exclusion chromatography (SEC), and IEC. Consistent among these examples are the ability to employ high mobile phase flow rates with minimal back pressures. The most promising applications are in the separation of macromolecules, where the mass transfer characteristics of the phases result in the ability to run at high linear velocities without penalty. This bodes well in particular for process applications.

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“…Furthermore, it was observed that the overloading effects appeared on different types of columns with C18 ligands, independent of the base matrix (silica, hybrid silica, or polymeric). Additionally, overloading effects are also found with non‐ligated polymeric phases, which do not contain C18 ligands or silanols . However, results showed that peak tailing, which appears on traditional C18 stationary phases under conditions of sample volume overload, were not observed on nonderivatized polymeric phases.…”

Section: Introductionmentioning

confidence: 97%

Overload Effects in Reversed Phase Protein Separations using Capillary‐Channeled Polymer Fiber Columns

Wang

Marcus

2018

Biotechnology Progress

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In preparative chromatography, overloading effects are a fact of life. However, peak distortion is problematic in analytical scale chromatography, especially in the case of bio-macromolecule separations. Capillary-channeled polymer fibers have been employed for fast protein separations in reversed phase, ion exchange, and hydrophobic interaction liquid chromatography. Although the primary advantage of the phase is operation at high linear velocities (~100 mm s ) without van Deemter C-term limitations, the limited specific surface area (<5 m g ) suggests that a thorough understanding of overloading effects is needed. We evaluate important factors (injected mass and volume) affecting overload in terms of peak height, width and shape (asymmetry). Overload conditions readily apparent as the peak shape changes from a Gaussian distribution to a left-triangle with slight tailing; more-or-less classical overload behavior. Three methods were used to compute column efficiency in terms of plate counts (N), including the area height, half-height, and the Dorsey-Foley approaches. The half-height method is best-suited for describing column overload, accounting for peak distortion and achieving high levels of consistency. The limiting plate count (N ) and the column sample loading capacity (ω ) were key metrics to characterize overall column performance. Peak capacity (P) was used to assess gradient elution separation performance. Increased flow rates and column length result in enhanced loading capacity. pH mismatch between the sample matrix and the mobile phase, a common cause of protein overload with other phases, was not responsible for distorted peak shapes unless the solvent pH is close to the pI of protein samples.

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Short fibrous supports for preparative chromatographic separations of biomolecules

Cited by 22 publications

References 20 publications

Initial evaluation of protein throughput and yield characteristics on nylon 6 capillary‐channeled polymer (C‐CP) fiber stationary phases by frontal analysis

Initial evaluation of protein throughput and yield characteristics on nylon 6 capillary‐channeled polymer (C‐CP) fiber stationary phases by frontal analysis

Use of polymer fiber stationary phases for liquid chromatography separations: Part II – applications

Overload Effects in Reversed Phase Protein Separations using Capillary‐Channeled Polymer Fiber Columns

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