Deconvolution of high‐throughput multicomponent isotherms using multivariate data analysis of protein spectra

Abstract

Gaining a more profound understanding of biopharmaceutical downstream processes is a key demand of the Quality by Design (QbD) guidelines. One of the most dominant approaches to gain process understanding is the extensive use of experimental high‐throughput formats, such as batch chromatography on robotic liquid handling stations. Using these high‐throughput experimental formats, the generation of numerous samples poses an enormous problem to subsequent analytical techniques. Here, a high‐throughput case study for batch chromatographic multicomponent isotherms is presented. To debottleneck the subsequent analytics, a noninvasive technique using UV spectra and multivariate statistics was adapted to a batch chromatographic format. Using this approach, it was possible to integrate the entire analytical setup into the robotic workflow. As a case study, batch isotherms for sulfopropyl sepharose fast flow and the model proteins cytochrome c and lysozyme at various pH values and ionic strengths were recorded. A successful examination of the quality of the analytical procedure compared to classical single wavelength photometry was carried out. To address the growing demand for a more profound process understanding, the experimental data were fitted to the steric mass action isotherm, getting a more detailed insight into the competitive binding behavior at various pH values and ionic strengths.

Automation and miniaturization: enabling tools for fast, high‐throughput process development in integrated continuous biomanufacturing

“…52 Furthermore, using the already established tools for more complex or precise analysis, such as precise determination of resin volume using optical methods in a 384-well plate, 53 or coupling the analytical tools with models to aid in the analysis of complex systems, where recent work highlights the use of such methodology for the study of complex systems as multicomponent isotherms in HTS platforms. [54][55][56] Scale-down models for different UO Another important aspect to consider is the feasibility of scaledown models in translating the results obtained at smaller scales into manufacturing scale processing. 21 Although some UO have favoured from a lot of attention from research peers, such as chromatography, some still lack feasible or practical scale-down models.…”

“…The incorporation of multi‐well plate readers into LHS and adaptation of the microfluidic device analytics, both on‐ and off‐chip, for accurate assays that have results in real time highlight the importance of having analytical tools that are adequate for the screening scale 52 . Furthermore, using the already established tools for more complex or precise analysis, such as precise determination of resin volume using optical methods in a 384‐well plate, 53 or coupling the analytical tools with models to aid in the analysis of complex systems, where recent work highlights the use of such methodology for the study of complex systems as multicomponent isotherms in HTS platforms 54‐56 …”

Downstream process development strategies for effective bioprocesses: Trends, progress, and combinatorial approaches

“…The plaques are generated by applying vacuum to adsorbent slurry, which is sucked into cavities of defined volume. The data generated using this device are of modeling quality as was shown for multi‐component isotherms of lysozyme and cytochrome c . Muthukumar and Rathore presented an MTP‐based membrane chromatography approach using AcroPrep Advance 96‐Well filter plates of 7 μL membrane volume (Pall Corp.) for the Granulocyte Colony‐Stimulating Factor, which is an alternative approach to chromatography based on adsorptive beads.…”

Cross‐scale quality assessment of a mechanistic cation exchange chromatography model

“…For mechanistic modeling of mAbs and other proteins in ion exchange chromatography, the SMA adsorption isotherm is frequently used in academic and industrial case studies. 16,[28][29][30][31][32][33] The SMA isotherm describes the multipoint binding of proteins to the resin under consideration of a protein's characteristic charge, the thermodynamic equilibrium of the adsorption process, and steric shielding effects. Multiple studies have demonstrated successful application of mechanistic models for the scale-up of chromatography processes.…”

“…They consist of partial differential equations, describing macroscopic transport through the column, mass transport within the stationary phase, and adsorption of protein to the resin. For mechanistic modeling of mAbs and other proteins in ion exchange chromatography, the SMA adsorption isotherm is frequently used in academic and industrial case studies 16,28‐33 . The SMA isotherm describes the multipoint binding of proteins to the resin under consideration of a protein's characteristic charge, the thermodynamic equilibrium of the adsorption process, and steric shielding effects.…”