Analytical strategies for the characterization of therapeutic monoclonal antibodies

Fekete, Szabolcs; Gassner, Anne‐Laure; Rudaz, Serge; Schappler, Julie; Guillarme, Davy

doi:10.1016/j.trac.2012.09.012

Cited by 112 publications

(87 citation statements)

References 41 publications

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“…During sample preparation, antibodies are denatured in the presence of an ionic detergent which masks the native charge of the protein providing a net negative charge enabling electrophoretic mobility separation based almost exclusively on the hydrodynamic radius of the protein molecule(s) [14]. The disulfide bonds which are responsible for the tetrameric structure of monoclonal antibodies are disrupted (reducing) or protected (non-reducing) during heat denaturation by reducing or alkylating reagents respectively [15,16].…”

Section: Introductionmentioning

confidence: 99%

Implementation of USP antibody standard for system suitability in capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for release and stability methods

Esterman

Katiyar

Krishnamurthy

2016

Journal of Pharmaceutical and Biomedical Analysis

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Section: Introductionmentioning

confidence: 99%

Implementation of USP antibody standard for system suitability in capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for release and stability methods

Esterman

Katiyar

Krishnamurthy

2016

Journal of Pharmaceutical and Biomedical Analysis

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“…Proteins and monoclonal antibodies (mAbs) are an emerging class of therapeutic agents currently being developed by many pharmaceutical companies [1]. Due to the increasing number of approved therapeutic proteins in the pharmaceutical area and the number of biosimilars (or follow-on-biologics) potentially entering the market, the need for analytical techniques for their detailed characterization has increased.…”

Section: Introductionmentioning

confidence: 99%

Ion-exchange chromatography for the characterization of biopharmaceuticals

Fekete

Beck

Veuthey

et al. 2015

Journal of Pharmaceutical and Biomedical Analysis

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“…[2][3][4][5][6][7][8][9][10][11] Several analytical methods have been developed for these purposes, for example, SDS polyacrylamide gel electrophoresis for the determination of the purity of proteins, size exclusion chromatography for aggregation, ion exchange chromatography or hydrophobic interaction chromatography for the detection of deamidation and oxidation variants, and Ellman's assay for quantification of free thiol groups. 12,13 Capillary-based SDS electrophoresis (CE-SDS) is one version of the capillary gel electrophoresis (CGE) using soluble linear polymers. Agarose and cross-linked polyacrylamide were often used as sieving matrices in the 1980s, 14,15 but the life time and reproducibility were poor.…”

Section: Introductionmentioning

confidence: 99%

Application of Microchip Electrophoresis Sodium Dodecyl Sulfate for the Evaluation of Change of Degradation Species of Therapeutic Antibodies in Stability Testing

et al. 2014

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We evaluated the performance of a commercial microchip electrophoresis instrument (LabChip ® GXII) for the evaluation of change of degradation species of therapeutic antibodies in stability testing. This system requires a sample volume of only 5 μg, and indicates fine resolution of size variant species such as light chain, heavy chain, non-glycosylated heavy chain and various degradation species. Precision and accuracy were high; the intermediate precision of 18 determinations was only 2.1% or less as RSD and recoveries ranged from 97.8 to 103.0% for major species as heavy chain, light chain and intact molecule of a therapeutic antibody. The applicability of this method was demonstrated by applying the method for the analysis of heat-degraded products of three pharmaceutical antibodies. Though some fragment peaks commonly appeared and increased according to temperature regardless of the source of preparations, one of them indicated specific peaks implying the cleavage of the peptide chain of the heavy chain. We also compared the performance of the method with those using conventional capillary-based SDS electrophoresis. Although the absolute purity values expressed as peak area % were different for the two methods, probably due to the difference in the detection methods, similar quality profiles were obtained within 40 s by microchip-based SDS electrophoresis. In addition, the degradation manner of three marketed antibodies depending on temperature was almost the same for the two methods. At the first stage in the development of manufacturing antibody pharmaceuticals, various factors including cell selection, cell cultivation, and formulation development should be evaluated using limited sample amounts. The stability testing using microchip-based electrophoresis seems suitable for these purposes.

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Analytical strategies for the characterization of therapeutic monoclonal antibodies

Cited by 112 publications

References 41 publications

Implementation of USP antibody standard for system suitability in capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for release and stability methods

Implementation of USP antibody standard for system suitability in capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for release and stability methods

Ion-exchange chromatography for the characterization of biopharmaceuticals

Application of Microchip Electrophoresis Sodium Dodecyl Sulfate for the Evaluation of Change of Degradation Species of Therapeutic Antibodies in Stability Testing

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