Application of the Steric Mass Action formalism for modeling under high loading conditions: Part 2. Investigation of high loading and column overloading effects

Seelinger, Felix; Wittkopp, Felix; Hirschheydt, Thomas von; Frech, Christian

doi:10.1016/j.chroma.2022.463266

Cited by 4 publications

(4 citation statements)

References 30 publications

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“…which is the basis of the self-association isotherm. [40,[54][55][56] However, the current interpretation was in conflict with the previously accepted explanation based on the same molecular weight.…”

Section: Determination Of Sma Parameterscontrasting

confidence: 97%

Standardized approach for accurate and reliable model development of ion‐exchange chromatography based on parameter‐by‐parameter method and consideration of extra‐column effects

Chen,

Lu,

Wang

et al. 2024

Biotechnology Journal

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Developing an accurate and reliable model for chromatographic separation that meets regulatory requirements and ensures consistency in model development remains challenging. In order to address this challenge, a standardized approach was proposed in this study with ion‐exchange chromatography (IEC). The approach includes the following steps: liquid flow identification, system and column‐specific parameters determination and validation, multi‐component system identification, protein amount validation, steric mass action parameters determination and evaluation, and validation of the calibrated model's generalization ability. The parameter‐by‐parameter (PbP) calibration method and the consideration of extra‐column effects were integrated to enhance the accuracy of the developed models. The experiments designed for implementing the PbP method (five gradient experiments for model calibration and one stepwise experiment for model validation) not only streamline the experimental workload but also ensure the extrapolation abilities of the model. The effectiveness of the standardized approach is successfully validated through an application about the IEC separation of industrial antibody variants, and satisfactory results were observed with R2 ≈ 0.9 for the majority of calibration and validation experiments. The standardized approach proposed in this work contributes significantly to improve the accuracy and reliability of the developed IEC models. Models developed using this standardized approach are ready to be applied to a broader range of industrial separation systems, and are likely find further applications in model‐assisted decision‐making of process development.

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Section: Determination Of Sma Parameterscontrasting

confidence: 97%

Standardized approach for accurate and reliable model development of ion‐exchange chromatography based on parameter‐by‐parameter method and consideration of extra‐column effects

Chen,

Lu,

Wang

et al. 2024

Biotechnology Journal

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“…This discrepancy might be attributed to the occurrence of multilayer adsorption at high loading density, and variants with protein–protein interaction might tend to elute earlier as a “preshoulder” (Seelinger et al, 2023). While the current model can only describe the case of single‐layer adsorption (Equation 2), other models like multistate adsorption model (Diedrich et al, 2017), multicomponent model (Seelinger et al, 2022), asymmetric activity coefficient (Huuk et al, 2017), hybrid model (Narayanan et al, 2021), and colloidal particle adsorption model (Briskot et al, 2021; Rischawy et al, 2022) can be utilized to address high loading density issues and further enhance the prediction accuracy. Although there was a discrepancy in the model prediction in the first half of elution profile, the collection started at the second half (retention time >40 min) with accurate model prediction (RMSE = 4.6%), resulting in a relatively minor impact on the collection window, yield and acidic variant content prediction.…”

Section: Resultsmentioning

confidence: 99%

“…This discrepancy might be attributed to the occurrence of multilayer adsorption at high loading density, and variants with protein-protein interaction might tend to elute earlier as a "preshoulder" (Seelinger et al, 2023). While the current model can only describe the case of single-layer adsorption (Equation 2), other models like multistate adsorption model (Diedrich et al, 2017), multicomponent model (Seelinger et al, 2022), asymmetric activity coefficient (Huuk et al, 2017), hybrid model (Narayanan et al, 2021), and colloidal particle adsorption model (Briskot et al, 2021;Rischawy et al, 2022) The analysis results indicated that when extending the well-fitted model parameters to higher loading density, a re-evaluation is necessary. The reassessment should not only encompass the model accuracy but, more importantly, the impact of the model's accuracy on the products attributes.…”

Section: Dt Test Under Extreme Conditionsmentioning

confidence: 92%

Realization of digital twin for dynamic control toward sample variation of ion exchange chromatography in antibody separation

Shi,

Chen,

Zhong

et al. 2024

Biotech & Bioengineering

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Digital twin (DT) is a virtual and digital representation of physical objects or processes. In this paper, this concept is applied to dynamic control of the collection window in the ion exchange chromatography (IEC) toward sample variations. A possible structure of a feedforward model‐based control DT system was proposed. Initially, a precise IEC mechanistic model was established through experiments, model fitting, and validation. The average root mean square error (RMSE) of fitting and validation was 8.1% and 7.4%, respectively. Then a model‐based gradient optimization was performed, resulting in a 70.0% yield with a remarkable 11.2% increase. Subsequently, the DT was established by systematically integrating the model, chromatography system, online high‐performance liquid chromatography, and a server computer. The DT was validated under varying load conditions. The results demonstrated that the DT could offer an accurate control with acidic variants proportion and yield difference of less than 2% compared to the offline analysis. The embedding mechanistic model also showed a positive predictive performance with an average RMSE of 11.7% during the DT test under >10% sample variation. Practical scenario tests indicated that tightening the control target could further enhance the DT robustness, achieving over 98% success rate with an average yield of 72.7%. The results demonstrated that the constructed DT could accurately mimic real‐world situations and perform an automated and flexible pooling in IEC. Additionally, a detailed methodology for applying DT was summarized.

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“…Based on this, step elution of different salt concentrations is chosen to elute the protein of interest at high purity and concentration. There are numerous studies for proteins with commercially available resins in literature [23][24][25][26][27]. Comparably, systematic studies for pDNA are rare [13].…”

Section: Introductionmentioning

confidence: 99%

Desorption of plasmid DNA from anion exchangers: Salt concentration at elution is independent of plasmid size and load

Beck

Biechele

Repik

et al. 2023

J of Separation Science

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Detailed studies on the sorption behavior of plasmids on anion exchangers are rare compared to proteins. In this study, we systematically compare the elution behavior of plasmid DNA on three common anion exchange resins using linear gradient and isocratic elution experiments. Two plasmids of different lengths, 8 and 20 kbp, were studied and their elution characteristics were compared to a green fluorescent protein. Using established methods for determining retention characteristics of biomolecules in ion exchange chromatography lead to remarkable results. In contrast to the green fluorescent protein, plasmid DNA consistently elutes at one characteristic salt concentration in linear gradient elution. This salt concentration was the same independent of plasmid size but differed slightly for different resins. The behavior is consistent also at preparative loadings of plasmid DNA. Thus, only a single linear gradient elution experiment is sufficient to design elution in a process scale capture step. At isocratic elution conditions, plasmid DNA elutes only above this characteristic concentration. Even at slightly lower concentrations most plasmids remain tightly bound. We hypothesize, that the desorption is accompanied by a conformational change leading to a reduced number of available negative charges for binding. This explanation is supported by structural analysis before and after elution.

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Application of the Steric Mass Action formalism for modeling under high loading conditions: Part 2. Investigation of high loading and column overloading effects

Cited by 4 publications

References 30 publications

Standardized approach for accurate and reliable model development of ion‐exchange chromatography based on parameter‐by‐parameter method and consideration of extra‐column effects

Standardized approach for accurate and reliable model development of ion‐exchange chromatography based on parameter‐by‐parameter method and consideration of extra‐column effects

Realization of digital twin for dynamic control toward sample variation of ion exchange chromatography in antibody separation

Desorption of plasmid DNA from anion exchangers: Salt concentration at elution is independent of plasmid size and load

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