Self-Interaction Chromatography of mAbs: Accurate Measurement of Dead Volumes

Hedberg, Sarah; Heng, Jerry Y. Y.; Liddell, John

doi:10.1007/s11095-015-1758-3

Cited by 8 publications

(4 citation statements)

References 45 publications

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“…Figure 4 suggests that the 1 L, 20 cm H ChromNeX can offer PvF performance similar to columns of diameter <1 cm (e.g., see Hansson, 2013). The 1 L devices had dead volumes of approximately 2%; much less than the >5% that might be expected in cylindrical columns of <1 cm diameter (Hedberg et al, 2015).…”

Section: Resultsmentioning

confidence: 93%

Performance of a new family of modular, bed‐supported, chromatography devices

Platteau

Stroehlein

Alstine

et al. 2023

Biotech & Bioengineering

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Prepacked chromatography columns and cassette filtration units offer many advantages in bioprocessing. These include reduced labor costs and processing times, ease of storage, and enhanced process flexibility. Rectangular formats are particularly attractive as they can be easily stacked and multiplexed together for continuous processing. Cylindrical chromatography beds have dominated bioprocessing even though their bed support and pressure‐flow performance vary with bed dimensions. This work presents the performance of novel, rhombohedral chromatography devices with internally supported beds. They are compatible with existing chromatography workstations and can be packed with any standard commercial resin. The devices offer pressure‐flow characteristics independent of container‐volume, simple multiplexing, and separation performance comparable to cylindrical columns. Their bi‐planar, internal bed support allows mechanically less‐rigid resins to be used at up to four times higher maximal linear velocities, and productivities approaching 200 g/L/h for affinity resins, compared to the 20 g/L/h typical of many column‐based devices. Three 5 L devices should allow processing of up to 3 kg of monoclonal antibody per hour.

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Section: Resultsmentioning

confidence: 93%

Performance of a new family of modular, bed‐supported, chromatography devices

Platteau

Stroehlein

Alstine

et al. 2023

Biotech & Bioengineering

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“…This retention volume analysis was also found to be more representative of the aggregation behavior of protein . The chromatographic retention factor ( k ′) can be calculated from the retention volume V P : where V 0 is the dead volume which was calculated by injecting protein on a column without interactions as outlined in Hedberg et al Finally, B 22 can be calculated using the following the equation: …”

Section: Experimental Proceduresmentioning

confidence: 99%

“…where V 0 is the dead volume which was calculated by injecting protein on a column without interactions as outlined in Hedberg et al 42 Finally, B 22 can be calculated using the following the equation:…”

Section: ■ Experimental Proceduresmentioning

confidence: 99%

Mapping the mAb Aggregation Propensity Using Self-Interaction Chromatography as a Screening Tool

et al. 2018

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The osmotic second virial coefficient ( B), which describes protein-protein molecular interactions in solution, was determined using self-interaction chromatography (SIC) for an IgG1-type mAb across a wide range of solution conditions. These data were compared with its time dependent aggregation behavior, as determined using size-exclusion chromatography (SEC), and its temperature dependent aggregation behavior using dynamic light scattering (DLS) over a four-week period (SEC) or overnight (DLS). DLS and SEC gave consistent data on aggregation behavior, which correlated well with experimental B trends across the wide pH (4-9) and NaCl concentration (0-1.0 M) ranges studied. The IgG aggregated at pH 4 for 0.5-1.0 M NaCl concentrations and for 0 M NaCl concentrations at pH 8. Best stability against aggregation was exhibited for the pH range from 5 to 8 at 0.8-1.0 M NaCl. SIC data were able to be classified within the one-day solution conditions for aggregation, which were not identified for 2-3 weeks in the accelerated SEC stability study. The ability of SIC to provide such data rapidly reflects the fundamentally thermodynamic nature of this parameter and of the aggregation process itself. Proteins with attractive protein-protein interactions and negative B coefficients in the range -3 to -6 clearly exhibit aggregation behavior, while B values in the range 0 to 2 showed good stability toward aggregation. SIC allows the rapid screening of solution conditions for which mAbs will exhibit stability to aggregation while requiring 90% less time and material compared with that required for a conventional SEC aggregation study.

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“…Therefore, a critical aspect in SIC revolves around the choice and use of the stationary phase resin as well as the solution conditions in terms of adsorption chemistry (Rakel et al 2013; Hedberg et al 2015). This is particularly important in the immobilisation of therapeutic proteins such as mAbs that may have high costs.…”

Section: Introductionmentioning

confidence: 99%

Improved adsorption reactions, kinetics and stability for model and therapeutic proteins immobilised on affinity resins

2019

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Protein adsorption on solid state media is important for the industrial affinity chromatography of biotherapeutics and for preparing materials for self-interaction chromatography where fundamental protein solution thermodynamic properties are measured. The adsorption of three model proteins (lysozyme, catalase and BSA) and two antibodies (a monoclonal and a polyclonal antibody) have been investigated on commercial affinity chromatography media with different surface functionalities (Formyl, Tresyl and Amino). Both the extent of protein immobilised (mg protein/ml media) and the reaction kinetics are reported for a range of reaction conditions, including pH, differing buffers as well as the presence of secondary reactants (glutaraldehyde, sodium cyanoborohydride, EDC and NHS). Compared to the reaction conditions recommended by manufacturers as well as those reported in previous published work, significant increases in the extent of protein immobilisation and reaction kinetics are reported here. The addition of glutaraldehyde or sodium cyanoborohydride was found to be especially effective even when not directly needed for the adsorption to happen. For mAb and pIgG, immobilisation levels of 50 and 31 mg of protein/ml of resin respectively were achieved, which are 100% or more than previously reported. Enhanced levels were achieved for lysozyme of 120 mg/ml with very rapid reaction kinetics (< 1 h) with sodium cyanoborohydride. It can be concluded that specific chromatography resins with Tresyl activated support offered enhanced levels of protein immobilisation due to their ability to react to form amine or thio-ether linkages with proteins. Additionally, glutaraldehyde can result in higher immobilisation levels whilst it can also accelerate immobilisation reaction kinetics.

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Self-Interaction Chromatography of mAbs: Accurate Measurement of Dead Volumes

Cited by 8 publications

References 45 publications

Performance of a new family of modular, bed‐supported, chromatography devices

Performance of a new family of modular, bed‐supported, chromatography devices

Mapping the mAb Aggregation Propensity Using Self-Interaction Chromatography as a Screening Tool

Improved adsorption reactions, kinetics and stability for model and therapeutic proteins immobilised on affinity resins

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