Ion-exchange chromatography for the characterization of biopharmaceuticals

Fekete, Szabolcs; Beck, Alain; Veuthey, Jean‐Luc; Guillarme, Davy

doi:10.1016/j.jpba.2015.02.037

Cited by 206 publications

(122 citation statements)

References 89 publications

(132 reference statements)

Supporting

Mentioning

117

Contrasting

Order By: Relevance

“…For optimal binding, in theory, the pH of the buffer has to be at least 1–2 pH units above the pI of the protein (Fekete et al 2015). At pH 8.7, the impurities should not bind, as they are weakly negatively charged, neutral, or even positively charged (pI between pH 9.5 and 7.0).…”

Section: Resultsmentioning

confidence: 99%

A novel scalable, robust downstream process for oncolytic rat parvovirus: isoelectric point-based elimination of empty particles

Leuchs

Frehtman

Riese

et al. 2017

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

The rodent protoparvovirus H-1PV, with its oncolytic and oncosuppressive properties, is a promising anticancer agent currently under testing in clinical trials. This explains the current demand for a scalable, good manufacturing practice-compatible virus purification process yielding high-grade pure infectious particles and overcoming the limitations of the current system based on density gradient centrifugation. We describe here a scalable process offering high purity and recovery. Taking advantage of the isoelectric point difference between full and empty particles, it eliminates most empty particles. Full particles have a significantly higher cationic charge than empty ones, with an isoelectric point of 5.8–6.2 versus 6.3 (as determined by isoelectric focusing and chromatofocusing). Thanks to this difference, infectious full particles can be separated from empty particles and most protein impurities by Convective interaction media® diethylaminoethyl (DEAE) anion exchange chromatography: applying unpurified H-1PV to the column in 0.15 M NaCl leaves, the former on the column and the latter in the flow through. The full particles are then recovered by elution with 0.25 M NaCl. The whole large-scale purification process involves filtration, single-step DEAE anion exchange chromatography, buffer exchange by cross-flow filtration, and final formulation in Visipaque/Ringer solution. It results in 98% contaminating protein removal and 96% empty particle elimination. The final infectious particle concentration reaches 3.5E10 plaque forming units (PFU)/ml, with a specific activity of 6.8E11 PFU/mg protein. Overall recovery is over 40%. The newly established method is suitable for use in commercial production.

show abstract

Section: Resultsmentioning

confidence: 99%

A novel scalable, robust downstream process for oncolytic rat parvovirus: isoelectric point-based elimination of empty particles

Leuchs

Frehtman

Riese

et al. 2017

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

show abstract

“…Also, the hydrophobic nature of PDMS can lead to the adsorption of biomolecules onto channel walls and flexibility of this elastomer limits pressures to just a few bar [15]. These constraints suggest that ion exchange chromatography methods that rely on pH and salt gradients in aqueous solutions [27,28,31] might be the most promising use of packed beds in PDMS, especially for short lengths used for solid phase extraction that require lower pressures [32]. …”

Section: Discussionmentioning

confidence: 99%

Advances in and prospects of microchip liquid chromatography

Grinias

Kennedy

2016

TrAC Trends in Analytical Chemistry

View full text Add to dashboard Cite

This review focuses on recent advances in the field of microfluidic liquid chromatography from January 2013 through April 2015. Articles are organized by the type of stationary phase support focusing on device fabrication, column preparation, and use for specific applications. Additionally, a comprehensive table comparing chromatographic figures of merit for the work described is included as Appendix A as a reference for readers.

show abstract

“…Hence, health authorities consider charge heterogeneity profiling of biopharmaceuticals as important for product characterization and product stability assessment. For this purpose, different analytical tools for charge heterogeneity profiling, including IEC , cIEF , and CZE are available. Since these techniques utilize different separation principles, their performance may depend on the individual characteristics of the investigated pharmaceutical and then requires a project specific method selection.…”

Section: Introductionmentioning

confidence: 99%

Application of affinity capillary electrophoresis for charge heterogeneity profiling of biopharmaceuticals

et al. 2019

View full text Add to dashboard Cite

Charge heterogeneity profiling is important for the quality control (QC) of biopharmaceuticals. Because of the increasing complexity of these therapeutic entities [1], the development of alternative analytical techniques is needed. In this work, flow‐through partial‐filling affinity capillary electrophoresis (FTPFACE) has been established as a method for the analysis of a mixture of two similar monoclonal antibodies (mAbs). The addition of a specific ligand results in the complexation of one mAb in the co‐formulation, thus changing its migration time in the electric field. This allows the characterization of the charged variants of the non‐shifted mAb without interferences. Adsorption of proteins to the inner capillary wall has been circumvented by rinsing with guanidine hydrochloride before each injection. The presented FTPFACE approach requires only very small amounts of ligands and provides complete comparability with a standard CZE of a single mAb.

show abstract

Ion-exchange chromatography for the characterization of biopharmaceuticals

Cited by 206 publications

References 89 publications

A novel scalable, robust downstream process for oncolytic rat parvovirus: isoelectric point-based elimination of empty particles

A novel scalable, robust downstream process for oncolytic rat parvovirus: isoelectric point-based elimination of empty particles

Advances in and prospects of microchip liquid chromatography

Application of affinity capillary electrophoresis for charge heterogeneity profiling of biopharmaceuticals

Contact Info

Product

Resources

About