Performance evaluation of an automated and continuous antibody purification process in a side-by-side comparability study

Continuous manufacturing (CM) introduces the benefits of cost efficiency, reliability and scalability for the manufacturing of biopharmaceuticals. Higher flexibility, smaller facility footprints and cost of goods benefits are advantages of this production mode. It offers high flexibility in regard of demand changes from clinical to launch and for volatile market dynamics. In combination with disposable equipment, faster time-to-market and closed processing seems feasible. Bayer´s unique CM platform consists of a series of downstream processing (DSP) unit operations through which the drug substance moves continuously and all unit operations happen more or less in parallel at the same time. The technology offers the potential to make Quality by Design (QbD) a reality (with continuously monitored process parameters and real-time feedback process control to maintain quality-indicating parameters within limits at all times, multi-variate data analysis). Individual unit operations are intelligently integrated and critical process parameters are monitored and controlled in real-time. Conditioning modules allow immediate corrective actions to be executed in an automated fashion to maintain the entire process in a state of control with low batch-to-batch variability. In addition, online sampling and testing functions provide early warning of potential excursions. By reduced manual interference this will also lead to reduction of operator errors and according deviations. Manufacturing facilities will be significantly less capital-intensive (e.g. by simpler layout) than large, traditional batch facilities as disposable technology and aseptic connections offer superior protection against bioburden ingress and other forms of contamination. The presentation also intends to illustrate comparability of CM versus batch processing in a side-by-side approach covering process information, real time analysis as well as quality data from intermediates and final drug substance of an antibody product. Please click Additional Files below to see the full abstract

Towards the implementation of a continuous bioprocess in single use technology

Bayer has developed a new production technology for monoclonal antibodies based on single use equipment and continuous processing. The development is referred to as the MoBiDiK project (Modular, Biologics, Disposable and Continuous) and was presented at the ICB 1 and ICB 2 conferences. At the ICB 3 conference we will focus on the topics that are critical to the implementation of the technology in a GMP production environment. These are: Process robustness, GMP readiness of the equipment, automation as well as process control and product release strategies

Pilot-scale process development for the purification of the recombinant antibody 2G12 from transgenic tobacco

The production of recombinant therapeutic proteins is becoming increasingly important as demand rises and the industry reaches its production capacity. Transgenic plants offer an alternative production system to the established platforms, but this requires the development of downstream processing schemes tailored for both the platform and the product. The aim of this thesis, within the EU-funded project Pharma-Planta, was to develop and optimize downstream processing strategies for the purification of the recombinant anti-HIV antibody 2G12. Different techniques were successfully combined into a process strategy to fulfill this objective. Clinical-grade recombinant antibody was purified using by the following process steps and industrially available equipment: manual harvest, dispersion and extraction, pH shift in the crude extract, fiber removal by filtration, clarification by filtration, capture by affinity Protein A membrane chromatography, intermediate purification by ceramic hydroxyapatite chromatography, polishing by virus filtration, concentration by ultrafiltration, diafiltration by ultrafiltration, final concentration and formulation. The process recovery of 60% resulted in a final yield of 3.7 g of clinical-grade tobacco-derived antibody purified from 216 kg leaf material, equal to 880 l crude extract. The antibody was polished to a purity >99%, consisting of 99.9% monomeric Ab with an in vitro binding activity of 93% and an in vitro neutralizing activity of 103% relative to the CHO2G12. This thesis established the basis for reproducible and predictable processing of plant material at the hundred-kilogram scale, and shows how further upscaling could be achieved

Pilot-scale process development for the purification of the recombinant antibody 2G12 from transgenic tobacco

The production of recombinant therapeutic proteins is becoming increasingly important as demand rises and the industry reaches its production capacity. Transgenic plants offer an alternative production system to the established platforms, but this requires the development of downstream processing schemes tailored for both the platform and the product. The aim of this thesis, within the EU-funded project Pharma-Planta, was to develop and optimize downstream processing strategies for the purification of the recombinant anti-HIV antibody 2G12. Different techniques were successfully combined into a process strategy to fulfill this objective. Clinical-grade recombinant antibody was purified using by the following process steps and industrially available equipment: manual harvest, dispersion and extraction, pH shift in the crude extract, fiber removal by filtration, clarification by filtration, capture by affinity Protein A membrane chromatography, intermediate purification by ceramic hydroxyapatite chromatography, polishing by virus filtration, concentration by ultrafiltration, diafiltration by ultrafiltration, final concentration and formulation. The process recovery of 60% resulted in a final yield of 3.7 g of clinical-grade tobacco-derived antibody purified from 216 kg leaf material, equal to 880 l crude extract. The antibody was polished to a purity >99%, consisting of 99.9% monomeric Ab with an in vitro binding activity of 93% and an in vitro neutralizing activity of 103% relative to the CHO2G12. This thesis established the basis for reproducible and predictable processing of plant material at the hundred-kilogram scale, and shows how further upscaling could be achieved

Reviewing the process intensification landscape through the introduction of a novel, multitiered classification for downstream processing

Abstract A demand for process intensification in biomanufacturing has increased over the past decade due to the ever‐expanding market for biopharmaceuticals. This is largely driven by factors such as a surge in biosimilars as patents expire, an aging population, and a rise in chronic diseases. With these market demands, pressure upon biomanufacturers to produce quality products with rapid turnaround escalates proportionally. Process intensification in biomanufacturing has been well received and accepted across industry based on the demonstration of its benefits of improved productivity and efficiency, while also reducing the cost of goods. However, while these benefits have been shown empirically, the challenges of adopting process intensification into industry remain, from smaller independent start‐up to big pharma. Traditionally, moving from batch to a process intensification scheme has been viewed as an “all or nothing” approach involving continuous bioprocessing, in which the factors of complexity and significant capital costs hinder its adoption. In addition, the literature is crowded with a variety of terms used to describe process intensification (continuous, periodic counter‐current, connected, intensified, steady‐state, etc.). Often, these terms are used inappropriately or as synonyms, which generates confusion in the field. Through a detailed review of current state‐of‐the‐art systems, consumables, and process intensification case studies, we herein propose a defined approach in the implementation of downstream process intensification through a standardized nomenclature and viewing it as distinct independent levels. These can function separately as intensified single‐unit operations or be built upon by integration with other process steps allowing for simple, incremental, cost‐effective implementation of process intensification in the manufacturing of biopharmaceuticals