Model‐based high‐throughput process development for chromatographic whey proteins separation

Abstract

In this study, an integrated approach involving the combined use of high-throughput screening (HTS) and column modeling during process development was applied to an industrial case involving the evaluation of four anion-exchange chromatography (AEX) resins and four hydrophobic interaction chromatography (HIC) resins for the separation of whey proteins having close pIs. From the HTS data, one resin of each type was selected (Capto Q and Octyl Sepharose 4 FF). Next, batch uptake experiments were performed to determine the adsorption isotherms of the major whey proteins on the selected resins, followed by isotherm parameters regression. Using the obtained isotherm parameters, the candidate chromatographic operations were modeled and experimentally validated. Finally, these were model-optimized and evaluated based on their optimized performances. In this example, Capto Q performed much better than Octyl Sepharose 4 FF in terms of column capacity, loading, throughput and productivity; hence, Capto Q was selected as the resin of choice for whey proteins separation. This operation was further scaled up from the lab scale (1 mL) to a preparative scale (35 L), with reproducible column elution profile. By this approach, a much wider space of operating variables could be investigated, thereby increasing the chances of finding the ideal operating conditions while keeping experimentation to the minimum.

Improved virus purification processes for vaccines and gene therapy

“…HTPS technologies are slowly becoming an established method in academic settings for improving process design (Nfor et al, ; Nestola et al, ; Osberghaus et al, ). Companies such as Tecan or Beckman Coulter—industrial leaders in automation and robotic platforms—are already developing fully customizable solutions for big pharma, SMEs, or small academic groups.…”

3D‐liquid chromatography as a complex mixture characterization tool for knowledge‐based downstream process development

“…For development of ion‐exchange operations the elution‐pH serves as a more reliable replacement parameter for the pI . Such small differences are regarded as a challenging separation problems, but systematic model‐based optimization of the process conditions has been shown to be capable of identifying conditions leading to sufficient process performance for a similar separation challenge …”

Separation of recombinant antibodies from DNA using divalent cations

“…High-throughput methods enable the rapid screening of suitable operation parameters [23][24][25][26]; although these methods are more popular for establishing chromatography methods, for nonchromatography methods it is simple to use high-throughput microtiter-plate technology to find suitable conditions. High-throughput methods enable the rapid screening of suitable operation parameters [23][24][25][26]; although these methods are more popular for establishing chromatography methods, for nonchromatography methods it is simple to use high-throughput microtiter-plate technology to find suitable conditions.…”

“…In the present investigation, our most striking observation was that conditions could be found for a CaCl 2 precipitation to reduce DNA levels and improve HMWI reduction while remaining compatible with other chromatographic and nonchromatographic purification steps. High-throughput methods enable the rapid screening of suitable operation parameters [23][24][25][26]; although these methods are more popular for establishing chromatography methods, for nonchromatography methods it is simple to use high-throughput microtiter-plate technology to find suitable conditions. Factorial experiment design guided us to the most important factors in our investigation: the use of CaCl 2 and its concentration.…”