Optimizing Downstream Processes for Biotherapeutic Development with Prepacked Chromatography Columns

Column cromatography | Image credit: ©Alen – stock.adobe.com

The growing demand for targeted drugs and biologics across numerous biomedical areas has driven the rapid development and approval of innovative biotherapeutics. For example, since 2015, the number of biologics approved by FDA has reached two-digit figures annually, compared with fewer than 10 per year previously (1). This shift reflects the industry’s notable progress in accelerating the development of novel drugs while underscoring the need for enhanced downstream production processes to maintain product efficacy, safety, and quality.

Biotherapeutic purification and production bottlenecks

Downstream processing during manufacturing of biotherapeutics involves recovering and purifying target molecules from a mixture containing host cell proteins, debris, and by-products of cell cultures. Chromatography is a technique widely used during this process for selective isolation and purification of target molecules and removal of impurities (2). However, as precision medicine necessitates biotherapeutics that are increasingly diverse and complex, significant challenges arise in downstream processing, such as higher production costs due to the intricacies of performing multiple isolation and purification steps. These challenges can negatively affect data reproducibility and create production bottlenecks, as well as compliance issues with regulatory standards. Therefore, chromatography workflows must be targeted to the unique physicochemical properties of each biomolecule to effectively remove impurities, such as DNA, host cell proteins, endotoxins, viruses, and protein aggregates, while also increasing process efficiencies.

Lead time for a product must take into account the labor required for column preparation, which includes determining the appropriate resin slurry concentration for a packed bed volume, column packing, testing, and qualification. Traditional column packing methods can also be complex, and irregularities can lead to uneven flow and cause band broadening, zone mixing, and flow rate variability, which reduce product yield and quality (3). Additionally, traditional systems can incur operational costs associated with cleaning, sterilization, and revalidation of equipment (4).

Along with the higher production costs of multiple chromatography steps, process scale-up from laboratory to manufacturing levels introduces additional complexities, as differences in column dimensions, resin behavior, and flow rates can render optimized, small-scale protocols unsuitable for large-scale downstream processing (5). Scale-up also increases resource demands, including larger space requirements for equipment and storage, hardware availability, resin availability for larger or multiple columns, and user training and expertise. Obtaining resins from different suppliers at different stages of the manufacturing process may also introduce variability and lead to inconsistent results, as individual suppliers provide distinct recommendations for their products. Understanding the nuances of slurry and buffer preparation, resin packing, and column cleaning and reuse for different processes requires technical expertise to ensure consistent performance. Overall, addressing these challenges is crucial to enabling successful large-scale manufacturing that meets stringent regulatory standards of purity and quality while ensuring efficient production and market entry.

Addressing downstream processing challenges with prepacked chromatography columns

Benefits in biotherapeutics manufacturing. The numerous challenges presented by in-house resin packing can be addressed with the use of prepacked chromatography columns (6). These columns are supplied ready-to-use by vendors—meeting good manufacturing practice (GMP) standards—and are accompanied by comprehensive documentation and usage instructions. Vendors offer columns in various sizes, prefilled with different resins and equipped with compatible hardware for a range of chromatography systems. Prepacked columns are expected to match or surpass the performance of in-house packed columns and meet the specifications required for scaling to larger manufacturing processes.

Prepacked columns used in biopharmaceutical manufacturing must comply with standards set by regulatory bodies to ensure the efficacy, quality, and safety of the final products. These regulations vary by country and the specific application of the columns. In the United States, FDA mandates that biopharmaceutical manufacturing adhere to current GMP (CGMP) regulations, ensuring that standardized processes are followed in controlled production environments (7). Additionally, FDA requires rigorous testing and validation to demonstrate the quality and purity of biopharmaceutical products. Prepacked chromatography columns used for the purification of these products must be validated for their packing consistency, performance reliability, and target molecule compatibility by the end user.

GMP-ready prepacked columns are available in various diameters, bed heights, and resin types (8). These columns support downstream processing across different stages of biopharmaceutical development and manufacturing, including purification and polishing chromatography steps for biomolecules, such as antibodies, recombinant proteins, or vaccines.

Overall, prepacked columns reduce the possibility of errors associated with in-house packing, such as variability between columns; thus, they enhance product yields and reproducibility of results. Moreover, the potential for prepacked columns to save time is a major draw for their application in the biopharmaceutical industry (9). Eliminating the need for column packing, validation, and cleaning can improve resource utilization. Owing to their plug-and-play design, prepacked columns minimize downtime and reduce requirements for specialized expertise or equipment, making them particularly advantageous for smaller facilities (6). Given that these columns are manufactured under controlled conditions, contamination risks are minimized during process optimization as well as between production batches, which is ideal in biopharmaceutical manufacturing. Use of prepacked columns avoids the introduction of environmental variables and potential cross-contamination, which can be an issue when columns are cleaned by hand and column hardware must be reused for multistep processes or different production batches.

Operational benefits. Prepacked chromatography columns can offer significant cost savings compared with traditional chromatography processes by eliminating manual column packing and validation, thereby lowering expenses related to materials and labor. Accordingly, there is no requirement for designated column packing areas, allowing production facility space to be used for other purposes and enabling simpler suite designs, thus improving operational efficiency. Further, keeping an inventory of prepacked columns may also reduce labor costs associated with resin management and handling.

Prepacked columns also provide increased flexibility via rapid process turnover, which may be advantageous for facilities producing multiple biotherapeutics. Process developers may also be able to choose from a wider range of resins during optimization and production of multiple biotherapeutics without specialized knowledge. Many resins are available in prepacked formats, allowing developers to adopt new separation technologies more easily and move beyond the constraints of “platform” resin strategies common in the industry (10).

Consistency is another key advantage offered by prepacked columns, as they are manufactured at standardized bed heights and diameters, ensuring optimized and consistent packing conditions. Certain columns are also manufactured with single lot resins. In addition, columns undergo pre-sanitization to minimize bioburden and avoid cross-contamination, are individually tested, and are provided to users with accompanying quality documentation. These measures may simplify processes and reduce labor requirements for quality control, with the potential to streamline regulatory approvals. Consistency is also an important consideration across facilities when biotherapeutics manufacturing occurs at multiple sites.

While an alternative to in-house column packing is to outsource the process to a column packing service, this may still involve determining resin availability, matching resin lots, and managing inventories if it is necessary for the customer to purchase and ship the resin to the service provider. Procurement of prepacked columns from a resin supplier that offers packing services simplifies the ordering process and requires contact with a single vendor that handles both resin inventory and technical support. This allows for streamlined communication and reduces delays by eliminating reliance on additional suppliers. Further, it may ensure reliability of production timelines by avoiding uncertainties in resin lead times and possible inconsistencies in column packing.

Adoption of prepacked columns in biopharmaceutical manufacturing. Prepacked columns have gained widespread use in downstream process development and small-scale studies, and they are becoming increasingly common in large-scale biopharmaceutical manufacturing operations that abide by GMP standards. The scalability of prepacked columns from small bench-scale to columns up to an inner diameter of 45 cm has been demonstrated (11). By reducing materials usage and eliminating the space and labor requirements for in-house column packing, as well as the subsequent need for performance testing and validation, prepacked columns can improve process efficiency. These improvements could lead to reduced overall costs in biopharmaceutical manufacturing, increased flexibility in production schedules, and rapid drug development. These factors could also potentially benefit patients by translating to reduced drug prices and providing faster access to necessary treatments.

A transformative solution

Prepacked chromatography columns have become a transformative solution in biopharmaceutical manufacturing by providing consistent performance and reproducibility, allowing developers to adhere to stringent quality and safety standards. This technology has been widely adopted by innovators and is becoming more common in many large-scale commercial environments, enabling greater operational efficiency, cost reduction, and quality improvement. With the continual need for rapid development of novel biotherapeutics, the use of prepacked columns will likely expand in downstream process optimization and production, driving industry innovations.

References

1. Martins, A.C.; Oshiro, M.Y.; Albericio, F.; de la Torre, B.G.; Pereira, G.J.V.; Gonzaga, R.V. Trends and Perspectives of Biological Drug Approvals by the FDA: A Review from 2015 to 2021. Biomedicines. 2022;10(9).
2. Rathore, A.S.; Kumar, D.; Kateja, N. Recent Developments in Chromatographic Purification of Biopharmaceuticals. Biotechnol Lett.2018 40(6):895-905.
3. Bemberis, I.; Noyes, A.; Natarajan, V. Column Packing for Process-Scale Chromatography: Guidelines for Reproducibility. BioPharm International. 2003 Supplement (2):23-30.
4. Rader, R.A., Langer, E. Single-Use Bioprocessing Equipment Trends and Adoption by CMOs. BioPharm International. 2018 31 (11):10-3.
5. Prentice, J.; Evans, S.T.; Robbins, D.; Ferreira, G. Pressure-Flow Experiments, Packing, and Modeling for Scale-up of a Mixed Mode Chromatography Column for Biopharmaceutical Manufacturing. J Chromatogr A. 2020 1625:461117.
6. Langer, E. Innovation in Pre-packed Disposable Chromatography Columns. BioPharm International. Single Use Systems ebook. 2014.
7. FDA. Current Good Manufacturing Practice (CGMP) Regulations. FDA.gov. 2023 (accessed June 25, 2025). https://www.fda.gov/drugs/pharmaceutical-quality-resources/current-good-manufacturing-practice-cgmp-regulations
8. Alsop, R.; Koley, S.; Jones, H.; Synder, M. Column Chromatography Scalability Study for a 33 cm Prepacked Foresight Pro Column from a 1.1 cm Bench Column. Presented at: ACS BIOT Annual Meeting, San Francisco, CA. August 13-17, 2023.
9. Scharl, T.; et al. Trend Analysis of Performance Parameters of Pre-packed Columns for Protein Chromatography Over a Time Span of Ten Years. J Chromatogr A. 2016 1465:63-70.
10. Allen, L. The Evolution of Platform Technologies for the Downstream Processing of Antibodies. In: Gottschalk U, editor. Process Scale Purification of Antibodies. Second ed: John Wiley & Sons, Inc.; 2017. p. 365-89.
11. Sidhu, G.; Exton, C.; Kronbetter, L.; Verona, E. Scalable Purification with Prepacked Ceramic Hydroxyapatite Process-scale Chromatography Columns. Presented at: ACS Spring 2025 Meeting; San Diego, CA. March 23-27, 2025. https://acs.digitellinc.com/p/s/scalable-purification-with-prepacked-ceramic-hydroxyapatite-process-scale-chromatography-columns-615783.

About the author

Laura Kronbetter is a Senior Global Product Manager at Bio-Rad Laboratories. She is a dedicated marketing professional in the life sciences industries with focused knowledge in managing and supporting process chromatography products used for the purification of biomolecules. With more than 20 years of experience in the industry and product management, Laura is passionate about developing and launching innovative products that meet customers' needs. She holds a bachelor's degree in biochemistry from Purdue University.