Determination of isoelectric points and relative charge variants of 23 therapeutic monoclonal antibodies

Goyon, Alexandre; Excoffier, Mélissa; Janin-Bussat, Marie Claire; Bobály, Balázs; Fekete, Szabolcs; Guillarme, Davy; Beck, Alain

doi:10.1016/j.jchromb.2017.09.033

Cited by 167 publications

(111 citation statements)

References 29 publications

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“…Commercially, mAb sales have exponentially increased since 2000, when the first mAb therapeutic was approved . In 2017, the USA Food and Drug Administration approved 68 mAbs for different therapeutic uses and more than 50 are currently under clinical trials, in the last evaluation phase .…”

Section: Introductionmentioning

confidence: 99%

Protein A chromatography: Challenges and progress in the purification of monoclonal antibodies

Ramos‐de‐la‐Peña

González‐Valdez

Aguilar

2019

J of Separation Science

112

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Antibodies for therapeutic use are being continuously approved and their demand has been steadily growing. As known, the golden standard for monoclonal antibody (mAb) purification is Protein A affinity chromatography, a technology that has gained high interest because of its great performance and capabilities. The main concerns are the elevated resins costs and their limited lifetime compared to other resins (e.g. ion exchange chromatography). Great efforts have been carried out to improve purification conditions, such as resin characterization and designing alkali/acid stable resins with a longer lifetime. Modification of Protein A ligands and alternative formats such as monoliths membranes and microshperes have been tested to increase the purification performance. New technology has been proposed to improve the large‐scale separation; in addition, alternative ligands have been suggested to capture mAbs instead of Protein A ligand; however, most of the information is locked by pharmaceutical companies. This mini review summarizes and describes the advances, results, and impact on the Protein A chromatography purification processing.

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Section: Introductionmentioning

confidence: 99%

Protein A chromatography: Challenges and progress in the purification of monoclonal antibodies

Ramos‐de‐la‐Peña

González‐Valdez

Aguilar

2019

J of Separation Science

112

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“…No trend was observed between both techniques regarding the percentage of basic variants ( R 2 = 0.30). The acidic variants were systematically higher in CIEF, but the generic method might overestimated the amount of acidic variants as discussed in . The choice of the data analysis software and integration parameters can also explain the differences in some cases but unlikely those observed in the present CZE study versus as the same operator manually integrated the peaks.…”

Section: Resultsmentioning

confidence: 70%

“…4A and 4B) and cation exchange chromatography (CEX) (Supporting Information Fig. 4C and 4D) from a previous study . Interestingly, 15 mAbs had higher amounts of basic variants with CEX performed in the salt elution mode, whereas 16 mAbs had lower amounts of acidic variants.…”

Section: Resultsmentioning

confidence: 86%

“…An increase of BGE pH from 5.7 to 7.0 is expected to improve the separation of charge variants for basic mAbs (p I > 8) but also increases the risk of protein adsorption to the capillary wall. The successful separations obtained with the developed CZE method for the most basic mAbs, i.e., ipilimumab, palivizumab and rituximab , account for a better dynamic coating obtained with 1.0% PEO compared to 0.05% HPMC. Indeed, detrimental effects were observed by He et al .…”

Section: Resultsmentioning

confidence: 92%

“…Similarly, CEX and HIC analysis have been performed to separate the two parental mAbs from a bi‐specific antibody in order to determine the heterodimerization efficiency . CE‐based techniques may gain more popularity for the separation of mAb mixtures, especially when antibodies are strongly adsorbed on the LC column stationary phase as for belimumab and ixekizumab in CEX .…”

Section: Resultsmentioning

confidence: 99%

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High‐resolution separation of monoclonal antibodies mixtures and their charge variants by an alternative and generic CZE method

et al. 2018

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The determination of mAb critical quality attributes (CQA) is crucial for their successful application in health diseases. A generic CZE method was developed for the high-resolution separation of various mAb charge variants, which are often recognized as important CQA. A dynamic coating of the capillary was obtained with polyethylene oxide (PEO), whereas Bis-Tris allowed the analysis of mAbs under native conditions at pH 7.0. The effect of PEO and Bis-Tris concentrations, as well as the nature of the acidic counter ion on the method performance was systematically studied. The %RSD on migration times was below 5% on three different CE instruments using the optimized method. Additional charge variants (in particular acidic variants) were resolved for 10 out of 17 mAbs compared to a reference CZE approach involving the use of ε-amino-caproic acid (EACA), triethylenetetramine (TETA), and hydroxypropylmethyl cellulose (HPMC). The amount of basic and acidic charge variants of 17 Food and Drug Administration (FDA) approved mAbs covering a wide range of physico-chemical properties, e.g., pI between 8.0 and 9.4 and different hydrophobicity, were mainly comprised between 5-15% and 15-30%, respectively. It is noteworthy that applications for the quality control in hospitals as well as for the combination of the immune checkpoint inhibitors nivolumab and ipilimumab were presented.

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Mass Spectrometry‐Based Strategies for Therapeutic Antibodies Extensive Characterization and Optimization (OptimAbs)

Bœuf

Debaene

Ayoub

et al. 2020

Structural Biology in Drug Discovery

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Determination of isoelectric points and relative charge variants of 23 therapeutic monoclonal antibodies

Cited by 167 publications

References 29 publications

Protein A chromatography: Challenges and progress in the purification of monoclonal antibodies

Protein A chromatography: Challenges and progress in the purification of monoclonal antibodies

High‐resolution separation of monoclonal antibodies mixtures and their charge variants by an alternative and generic CZE method

Mass Spectrometry‐Based Strategies for Therapeutic Antibodies Extensive Characterization and Optimization (OptimAbs)

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