Systematic method development in hydrophobic interaction chromatography I. Characterization of the phase system and modelling retention

Rippel, Géza; Bede, Á.; Szepesy, L.

doi:10.1016/0021-9673(94)01031-9

Cited by 13 publications

(11 citation statements)

References 32 publications

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“…It was experimentally shown by Szepesy and Karger that the retention of some common proteins in HIC gradient mode is in quantitative agreement with the LSS model [34,42]. Then, this model should enable retention prediction and modeling in HIC [43,44].…”

Section: Linear Solvent Strength Modelmentioning

confidence: 84%

“…Practical method development aspects are rarely reported in the literature for HIC [42]. In this section, the most important parameters affecting selectivity and retention in HIC are 8 described, and the possibility of automated method development is discussed.…”

Section: Methods Development In Hicmentioning

confidence: 99%

“…Szepesy and co-workers suggested, that the first step -for systematic method development in HIC -should be the selection of an appropriate phase system (the stationary and mobile phase) which is suitable for the required separation (provides adequate retention and selectivity) [42]. They developed a protocol for phase system characterization based on gradient experiments performed with three different gradient steepness performed on three columns and using three salts as mobile phase constituent.…”

Section: Computer Assisted Methods Optimization Retention Modellingmentioning

confidence: 99%

“…It can both increase and decrease the retention of hydrophobic and hydrophilic proteins, respectively [53,54]. The above results also imply that the effect of a salt cannot be predicted in advance but should always be determined experimentally as an early step of method development [42].…”

Section: Impact Of the Salt Nature And Concentrationmentioning

confidence: 96%

“…On the other hand, peak widths also vary with salt concentration, since it impacts the concentration gradient (steepness) and therefore the gradient band focusing effect. A systematic study showed the possibility to combine different salts (binary and ternary salt systems) to modify selectivity and retention in HIC [42]. In this phase system optimization concept, the experimental design was based on gradient experiments performed on three different columns and with three different buffers (salts).…”

Section: Impact Of the Salt Nature And Concentrationmentioning

confidence: 99%

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

Fekete

Veuthey

Beck³

et al. 2016

Journal of Pharmaceutical and Biomedical Analysis

122

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Hydrophobic interaction chromatography (HIC) is a historical strategy used for the analytical purification and characterization of proteins. Similarly to what can be done in reversed-phase liquid chromatography (RPLC), HIC is able to separate protein species based on their hydrophobicity, but using different conditions. Compared to RPLC, the main benefit of HIC is its ability to perform separations under non denaturing conditions (i.e. physiological pH conditions, ambient mobile phase temperature and no need for organic solvents) and so an orthogonal method. The goal of this review is to provide a general overview of theoretical and practical aspects of modern HIC applied for the characterization of therapeutic protein biopharmaceuticals including monoclonal antibodies (mAbs), antibody drug conjugates (ADCs) and bispecific antibodies (bsAbs). Therefore, method development approaches, state-of-the-art column technology, applications and future perspectives are described and critically discussed.

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