Separation of Macromolecules by Ultrafiltration: Influence of Protein Adsorption, Protein-Protein Interactions, and Concentration Polarization

Ingham, Kenneth C.; Busby, Thomas F.; Sahleström, Ylva; Castino, F

doi:10.1007/978-1-4613-3162-9_9

Cited by 45 publications

(22 citation statements)

References 17 publications

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“…The sieving coefficient trends for lysozyme in the presence of BSA as a function of pH, as well as ionic strength, agree with previous diafiltration (Ingham et al, 1980) and deadend ultrafiltration (Iritani et al, 1997) experiments. It was reported that the sieving coefficient at neutral pH increased as a function of ionic strength, in agreement with our results.…”

Section: Comparison Of Lysozyme Sieving Behavior With Previous Worksupporting

confidence: 87%

“…In diafiltration, the lysozyme concentration in the stirred cell (c) as a function of time, which was calculated from the concentration of lysozyme in the filtrate samples by difference, can be described by (Ingham et al, 1980) …”

Section: Sieving Coefficient Analysismentioning

confidence: 99%

“…It has been observed frequently that although the filtration properties of two proteins that are dissimilar in size suggest that they are amenable to separation, the sieving behavior of the smaller protein in the presence of the larger one is surprisingly poor (Blatt et al, 1970;Ingham et al, 1980;Shettigar et al, 1982;Gurland et al, 1984;Higuchi et al, 1991). There are several possible reasons for the poor separation behavior (Nakatsuka and Michaels, 1992;Saksena and Zydney, 1994), including the membrane pore size and pore size distribution, protein adsorption inside or on the upper surface of the membrane, bulk mass transfer effects (i.e., concentration polarization), and protein -protein interactions.…”

Section: Introductionmentioning

confidence: 99%

“…Several studies have demonstrated that the solution pH can affect the separation of binary mixtures significantly at low ionic strength (< 0.15M) (Chaufer et al, 2000;Ghosh and Cui, 1998;Nakatsuka and Michaels, 1992;Saksena and Zydney, 1994;Sannier et al, 1996;van Eijndhoven et al, 1995), most likely by influencing protein -membrane electrostatic interactions. However, several studies at elevated ionic strength have also shown that the separation behavior of protein mixtures can vary significantly as a function of ionic strength and pH, which has been speculated to be due to protein -protein interactions (Ingham et al, 1980;Iritani et al, 1997). Specifically, it was observed that at low ionic strength and neutral pH, where BSA (pI 4.9) and lysozyme (pI 11) are oppositely charged, lysozyme (14.3 kD) transmission through a 30 kD molecular weight cut-off (MWCO) membrane was surprisingly low in the presence of BSA (67 kD).…”

Section: Introductionmentioning

confidence: 99%

“…However, the transmission was significantly enhanced when the ionic strength was increased. It was suggested that the enhancement of lysozyme transmission as a function of ionic strength was due to the reduction in the degree of BSA/lysozyme heteroassociation in solution (Ingham et al, 1980) or in the filter cake above the membrane (Iritani et al, 1997). However, due to the difficulties of measuring weak cross-association between protein molecules, the role of protein-protein interactions in selective filtration has not been elucidated in situations such as these.…”

Section: Introductionmentioning

confidence: 99%

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Correlation of diafiltration sieving behavior of lysozyme‐BSA mixtures with osmotic second virial cross‐coefficients

Tessier

Verruto

Sandler

et al. 2004

Biotech & Bioengineering

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The role of protein -protein interactions in membrane separations of protein mixtures remains incompletely understood, largely due to the difficulty of characterizing protein self-and, especially, cross-association. Recently, a novel technique, cross-interaction chromatography, has been developed to measure weak protein cross-association in terms of the osmotic second virial cross-coefficient. In this work the relationship between protein cross-association and the sieving behavior of lysozyme in the presence of BSA has been investigated. Sieving coefficients were measured using a stirred diafiltration cell over a range of pH and ionic strength, and a striking correlation between the lysozyme sieving and second virial cross-coefficients for BSA/lysozyme mixtures has been found: when the protein cross-interactions are most attractive (negative second virial cross-coefficient), the lysozyme sieving coefficients are lowest, and vice versa. The correlation between the sieving and second virial crosscoefficients may be due to the physically similar environments in the chromatography and filtration experiments since one protein is passed through a concentrated region of the second protein either immobilized on the column or accumulated at the membrane surface, and the migration rate of the mobile protein in both cases is influenced by protein cross-association. This study represents the first time that molecular interactions in binary mixtures have been related directly to filtration behavior, and may provide a useful approach to optimize the separation of other binary protein mixtures. B 2004 Wiley Periodicals, Inc.

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Section: Comparison Of Lysozyme Sieving Behavior With Previous Worksupporting

confidence: 87%

Section: Sieving Coefficient Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Correlation of diafiltration sieving behavior of lysozyme‐BSA mixtures with osmotic second virial cross‐coefficients

Tessier

Verruto

Sandler

et al. 2004

Biotech & Bioengineering

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High performance tangential flow filtration

Reis

Gadam²,

Frautschy

et al. 1997

Biotechnol. Bioeng.

161

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Conventional tangential flow filtration (TFF) has traditionally been limited to separation of solutes that differ by about ten‐fold in size. Wide pore‐size distributions, membrane fouling, and concentration polarization phenomena have commonly been cited as reasons for this limitation. The use of TFF in the biotechnology industry has therefore been restricted to cell‐protein, virus‐protein, and protein‐buffer separations. A multi‐disciplinary team with industrial and academic members was formed to overcome these limitations and enable protein‐protein separations using High Performance TFF (HPTFF) systems. Pore‐size distributions have been improved with the development of new membrane formulation and casting techniques. Membrane fouling has been controlled by operating in the transmembrane pressure‐dependent regime of the filtrate flux curve and by carefully controlling fluid dynamic start‐up conditions. Concentration polarization was exploited to enhance, rather than limit, the resolution of solutes. Concentration polarization has also been controlled by operating a co‐current filtrate stream that maintains transmembrane pressure constant along the length of the TFF module. High yields and purification factors were obtained even with small differences in protein sieving. IgG‐BSA and BSA monomer‐oligomer mixtures have successfully been separated with these systems. HPTFF technology provides a competitive purification tool to complement chromatographic processing of proteins. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 71–82, 1997.

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Cross flow filtration of RNA extracts by hollow fiber membrane

Fernandez

Romano

Otero

1992

Acta Biotechnologica

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The feasibility of using ultrafiltration to concentrate RNA was investigated. Ultrafiltration flux and solute retentivity were affected by membrane cut-off and inlet pressure. The RNA can be concentrated by selecting a hydraulically permeable membrane (PM 30) and then operating the system at a sufficiently high pressure (2.07. lo5 Nm-') and a high recirculation rate (150 cm-I). The use of this procedure was limited to a high-flux regime. However, a very high retentivity of macrosolute was achieved. The effect of transmembrane pressure on the apparent mass transfer coeficient in the ,,gel polarization model" was discussed and related to ultrafiltration flux.

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Separation of Macromolecules by Ultrafiltration: Influence of Protein Adsorption, Protein-Protein Interactions, and Concentration Polarization

Cited by 45 publications

References 17 publications

Correlation of diafiltration sieving behavior of lysozyme‐BSA mixtures with osmotic second virial cross‐coefficients

Correlation of diafiltration sieving behavior of lysozyme‐BSA mixtures with osmotic second virial cross‐coefficients

High performance tangential flow filtration

Cross flow filtration of RNA extracts by hollow fiber membrane

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