Hydrophobic interaction chromatography for the characterization of monoclonal antibodies and related products

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

doi:10.1016/j.jpba.2016.04.004

Cited by 122 publications

(76 citation statements)

References 85 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Recent review papers detailed HIC method development and also showed some useful applications [22][23][24]. The goal of the present review was to summarize the main points of these works and complete them with most recent results.…”

Section: Hydrophobic Interaction Chromatography (Hic)mentioning

confidence: 99%

Current possibilities of liquid chromatography for the characterization of antibody-drug conjugates

Bobály

Fleury-Souverain²,

Beck

et al. 2018

Journal of Pharmaceutical and Biomedical Analysis

Self Cite

View full text Add to dashboard Cite

Antibody Drug Conjugates (ADCs) are innovative biopharmaceuticals gaining increasing attention over the last two decades. The concept of ADCs lead to new therapy approaches in numerous oncological indications as well in infectious diseases. Currently, around 60 CECs are in clinical trials indicating the expanding importance of this class of protein therapeutics. ADCs show unprecedented intrinsic heterogeneity and address new quality attributes which have to be assessed. Liquid chromatography is one of the most frequently used analytical method for the characterization of ADCs. This review summarizes recent results in the chromatographic characterization of ADCs and supposed to provide a general overview on the possibilities and limitations of current approaches for the evaluation of drug load distribution, determination of average drug to antibody ratio (DAR), and for the analysis of process/storage related impurities. Hydrophobic interaction chromatography (HIC), reversed phase liquid chromatography (RPLC), size exclusion chromatography (SEC) and multidimensional separations are discussed focusing on the analysis of marketed ADCs. Fundamentals and aspects of method development are illustrated with applications for each technique. Future perspectives in hydrophilic interaction chromatography (HILIC), HIC, SEC and ion exchange chromatography (IEX) are also discussed.

show abstract

Section: Hydrophobic Interaction Chromatography (Hic)mentioning

confidence: 99%

Current possibilities of liquid chromatography for the characterization of antibody-drug conjugates

Bobály

Fleury-Souverain²,

Beck

et al. 2018

Journal of Pharmaceutical and Biomedical Analysis

Self Cite

View full text Add to dashboard Cite

show abstract

“…Unlike RP, HIC operates under non denaturing conditions (no organic solvents, physiological pH, and ambient mobile phase temperature) and has shown success in separating charge variant species (Boyd, Kaschak, & Yan, ; Ouellette, Chumsae, Clabbers, Radziejewski, & Correia, ; Valliere‐Douglass, Wallace, & Balland, ). Recent improvements in the stationary phase for HIC have focused on decreasing particle size to enhance the kinetic performance, which may allow for greater industrial scale adoption (Fekete, Veuthey, Beck, & Guillarme, ).…”

Section: Principles Of Charge Variants Analyticsmentioning

confidence: 99%

“…Complete inactivation (Rehder et al, 2008) Succinimide Asn, Asp May illicit immune response (Chen et al, 1996) C-terminal Lys/Arg Lys, Arg No significant impact on binding, PK, or half-life (Alt et al, 2016;Antes et al, 2007;Khawli et al, 2010;Lyubarskaya et al, 2006) Ouellette, Chumsae, Clabbers, Radziejewski, & Correia, 2013;Valliere-Douglass, Wallace, & Balland, 2008). Recent improvements in the stationary phase for HIC have focused on decreasing particle size to enhance the kinetic performance, which may allow for greater industrial scale adoption (Fekete, Veuthey, Beck, & Guillarme, 2016). Table 2 shows selected representative works for analyzing charge variant species and the techniques used in those studies.…”

Section: Isomerization Aspmentioning

confidence: 99%

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

Chung

Tian

Tan

et al. 2018

Biotech & Bioengineering

View full text Add to dashboard Cite

Controlling the charge profile of therapeutic protein is a critical challenge in the current quality-by-design (QbD) paradigm, throughout all phases of biologics process development (PD): cell line development, upstream cell culture, recovery process, downstream purification, and analytical characterization. Charge variant profiles may influence efficacy and/or lead to unintended side-effects. Thus, maintaining a consistent charge profile is of tremendous importance, and increasingly, researchers have focused efforts toward developing strategies to mitigate variability during cell culture and to improve separation and detection of charge variants. Current understanding of factors affecting charge variant formation during manufacturing remains inadequate, and sometimes, even substantial commitment of resources may still not fully achieve the desired or consistent profiles. As such, this review attempts to provide a comprehensive resource for the biologics community by summarizing the impact of charge variants and CQA management, analytical methods for charge variant detection, as well as strategies in downstream and upstream PD for controlling charge variant profiles.

show abstract

“…Rituximab (pI of ∼9.1) and panitumumab (pI of ∼6.7) are intended to represent basic and acidic mAbs, respectively [26]. Ipilimumab is hydrophobic, while denosumab can be considered as a relatively hydrophilic mAb [32]. NISTmAb is currently the only universal standard reference material (SRM) for monoclonal antibody therapeutics.…”

Section: Development Of a General Hilic Gradientmentioning

confidence: 99%

Analysis of recombinant monoclonal antibodies in hydrophilic interaction chromatography: A generic method development approach

Bobály

D’Atri

Beck³

et al. 2017

Journal of Pharmaceutical and Biomedical Analysis

Self Cite

View full text Add to dashboard Cite

a b s t r a c tHydrophilic interaction liquid chromatography (HILIC) is a well-established technique for the separation and analysis of small polar compounds. A recently introduced widepore stationary phase expanded HILIC applications to larger molecules, such as therapeutic proteins. In this paper, we present some generic HILIC conditions adapted for a wide range of FDA and EMA approved recombinant monoclonal antibody (mAb) species and for an antibody-drug conjugate (ADC). Seven approved mAbs possessing various isoelectric point (pI) and hydrophobicity as well as a cysteine conjugated ADC were used in this study. Samples were digested by IdeS enzyme and digests were further fragmented by chemical reduction. The resulting fragments were separated by HILIC. The main benefit of HILIC was the separation of polar variants (glycovariants) in a reasonable analysis time at the protein level, which is not feasible with other chromatographic modes. Three samples were selected and chromatographic conditions were further optimized to maximize resolution. A commercial software was used to build up retention models. Experimental and predicted chromatograms showed good agreement and the average error of retention time prediction was less than 2%. Recovery of various species and sample stability under the applied conditions were also discussed.

show abstract

Hydrophobic interaction chromatography for the characterization of monoclonal antibodies and related products

Cited by 122 publications

References 85 publications

Current possibilities of liquid chromatography for the characterization of antibody-drug conjugates

Current possibilities of liquid chromatography for the characterization of antibody-drug conjugates

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

Analysis of recombinant monoclonal antibodies in hydrophilic interaction chromatography: A generic method development approach

Contact Info

Product

Resources

About