Predictive scaling of fiber‐based protein A capture chromatography using mechanistic modeling

The most straightforward method to increase monoclonal antibody (mAb) product yield is to complete the purification process in less steps. Here, three different fiber chromatographic devices were implemented using a holistic approach to intensify the mAb purification process and increase yield. Fiber protein A (proA) chromatography was first investigated, but traditional depth filtration was not sufficient in reducing the contaminant load as the fiber proA device prematurely fouled. Further experimentation revealed that chromatin aggregates were the most likely reason for the fiber fouling. To reduce levels of chromatin aggregates, a chromatographic clarification device (CCD) was incorporated into the process, resulting in single‐stage clarification of harvested cell culture fluid and reduction of DNA levels. The CCD clarified pool was then successfully processed through the fiber proA device, fully realizing the productivity gains that the fiber technology offers. After the proA and viral inactivation neutralization (VIN) hold step, the purification process was further intensified using a novel single‐use fiber‐based polishing anion exchange (AEX) material that is capable of binding both soluble and insoluble contaminants. The three‐stage fiber chromatographic purification process was compared to a legacy five‐step process of dual‐stage depth filtration, bead‐based proA chromatography, post‐VIN depth filtration, and bead‐based AEX chromatography. The overall yield from the five‐step process was 60%, while the fiber chromatographic‐enabled intensified process had an overall yield of 70%. The impurity clearance of DNA and host cell protein (HCP) for both processes were within the regulatory specification (<100 ppm HCP, <1 ppb DNA). For the harvest of a 2000 L cell culture, the intensified process is expected to increase productivity by 2.5‐fold at clarification, 50‐fold at the proA step, and 1.6‐fold in polishing. Relative to the legacy process, the intensified process would reduce buffer use by 1088 L and decrease overall process product mass intensity by 12.6%.

Section: Cell Culture and Productionmentioning

confidence: 99%

“…For the buffer use calculation, the elution volume of the fiber proA device is expected to decrease by approximately 30% at a large scale (Hahn et al, 2023). This assumption was included in this analysis.…”

Section: Analytical Measurementsmentioning

confidence: 99%

Fiber chromatographic enabled process intensification increases monoclonal antibody product yield

Anderson,

Seto,

Chau

et al. 2023

“…future facility designs that embrace modular (Bertran & Babi, 2024) and continuous concepts (Konoike et al, 2024;Minervini et al, 2024), novel purification technologies from fluidized bed systems and 3D-printed monoliths (Herlevi et al, 2024;Pei et al, 2024;Saballus et al, 2024) through to new membrane and chromatography devices (Lavoie et al, 2024;Platteau et al, 2024), bioprocess development acceleration tools that embrace high throughput tools with novel sensors, data analytics, modeling, and control strategies (Andersson et al, 2024;Banerjee et al, 2024;Dürauer et al, 2024;Hahn et al, 2024;Kaufman et al, 2024;Keulen et al, 2024;Lorek et al, 2024;Sebastian et al, 2024), separation science and solutions for non-mAb proteins (Rodriguez et al, 2024;Roush et al, 2024) and emerging viral products (Kish et al, 2024;Konstantinidis et al, 2024;Lyle et al, 2024;Perry et al, 2024;Soni et al, 2024), perspectives on the current status and future outlook for downstream processing of biological products drawn from the conference roundtable discussions (Jungbauer et al, 2024).…”

mentioning

confidence: 99%

Special Issue on “Recovery of Biological Products: Past, Present, Future”

Farid,

Tugcu,

Staby

2024

Downstream processing is pivotal in the biotherapeutic production sector, which has evolved from traditional recombinant proteins to cutting-edge modalities such as novel engineered antibody formats and gene therapies. The recent pandemic has further diversified pipelines, with a significant increase in research and development for groundbreaking viral-vectored and RNA products. This evolution introduces unique purification challenges, underscoring the need for manufacturing approaches that are more flexible, agile, scalable, digitally integrated, supply chain resilient, and capable of faster market delivery. This Special Issue of Biotechnology and Bioengineering (B&B) compiles advances and future directions in downstream processing of biotherapeutics, highlighting contributions from the recent Recovery of Biological Products XIX Conference (RXIX) with its theme on "Past, Present, Future" (RXIX, Rome, Italy, July 10-15, 2022) (Figure 1) chaired by Suzanne Farid (UCL), Nihal Tugcu (Sanofi), and Arne Staby (Novo Nordisk). The issue reflects on

Current state of implementation of in silico tools in the biopharmaceutical industry—Proceedings of the 5th modeling workshop

Wittkopp,

Welsh,

Todd

et al. 2024

The fifth modeling workshop (5MW) was held in June 2023 at Favrholm, Denmark and sponsored by Recovery of Biological Products Conference Series. The goal of the workshop was to assemble modeling practitioners to review and discuss the current state, progress since the last fourth mini modeling workshop (4MMW), gaps and opportunities for development, deployment and maintenance of models in bioprocess applications. Areas of focus were four categories: biophysics and molecular modeling, mechanistic modeling, computational fluid dynamics (CFD) and plant modeling. Highlights of the workshop included significant advancements in biophysical/molecular modeling to novel protein constructs, mechanistic models for filtration and initial forays into modeling of multiphase systems using CFD for a bioreactor and mapped strategically to cell line selection/facility fit. A significant impediment to more fully quantitative and calibrated models for biophysics is the lack of large, anonymized datasets. A potential solution would be the use of specific descriptors in a database that would allow for detailed analyzes without sharing proprietary information. Another gap identified was the lack of a consistent framework for use of models that are included or support a regulatory filing beyond the high‐level guidance in ICH Q8–Q11. One perspective is that modeling can be viewed as a component or precursor of machine learning (ML) and artificial intelligence (AI). Another outcome was alignment on a key definition for “mechanistic modeling.” Feedback from participants was that there was progression in all of the fields of modeling within scope of the conference. Some areas (e.g., biophysics and molecular modeling) have opportunities for significant research investment to realize full impact. However, the need for ongoing research and development for all model types does not preclude the application to support process development, manufacturing and use in regulatory filings. Analogous to ML and AI, given the current state of the four modeling types, a prospective investment in educating inter‐disciplinary subject matter experts (e.g., data science, chromatography) is essential to advancing the modeling community.