Adoption of single-use technology to entire production schemes has only recently been realized after decades of having disposable products available. The use of buffer and small disposable bioreactor bags was implemented early, while more sophisticated or large unit operations employing single-use technology are only recently available to complete the “disposable facility dream.”

Likewise, the implementation of continuous production will begin in a stepwise fashion, with individual unit operations addressing the largest bottleneck or by adopting the simplest, lowest-risk technology to achieve early ROI and productivity gains.

Multicolumn continuous chromatography (MCC) at production scale has been an ambition of GMP facilities for some time. Implementation of MCC at large scale greatly increases productivity, particularly at the capture step where many production facilities are bottlenecked as a result of productivity gains in upstream fed-batch or perfusion systems.

Beyond Continuous Batch Production

MCC by itself substantially intensifies the capture process with productivity gains of nearly triple.1 The bioprocess systems group of LEWA-Nikkiso America has further enhanced the chromatography process by engineering a platform that not only provides a simple two-column method for continuous capture (a patented process licensed from ChromaCon), but also uses innovative engineering and end-user verification to combine four unit operations into a single skid. Scale up of 100-fold has been proven by a GMP producer of biologics1 (Figure 1).

A single EcoPrime Twin® LPLC System can perform:

• Batch—with a range from0.4 to 20 LPM
• Continuous-batch capture—patented CaptureSMB® two-column technology
• Sequential (Integrated) polishing—pool-less two-step chromatography
• Buffer inline dilution (BID)—on-board inline dilution of buffers and CIP solutions
• Single-use aseptic interface—permits use in minimally classified suite.

Figure 1. Successful scale-up of 100-fold on two-column continuous capture systems. Instruments shown are the ContiChrom CUBE benchtop and EcoPrime Twin LPLC GMP pilot.

Simple Yet Flexible Multifunction System

Continuous, multicolumn Protein A capture chromatography is shown to debottleneck the downstream purification of monoclonal antibodies. The advantages of this type of continuous process, as compared to traditional single-column capture, include improved resin capacity utilization, shorter processing times or reduction in resin volume, and reduction in buffer compensation, all while maintaining yields at >98%.

However, achieving these benefits comes at the expense of increased hardware complexity and investment. More complex equipment also increases risk of downtime and maintenance costs (Figure 2).

While systems and processes utilizing up to as many as eight or more columns have been reported, a simple two-column continuous system maximizes process performance with respect to resin capacity utilization, buffer consumption, and product concentration while minimizing hardware investment and risk.2 Fewer columns significantly reduces equipment complexity simply by reducing the number of valves, plumbing, and monitoring required. Simpler hardware also means simpler operation and automation—facilitating operator understanding, equipment qualification, and process validation.

Employing continuous capture chromatography for process intensification is a proven way to enhance productivity. Combining multiple functions within a single unit operation on a single system (such as inline dilution of buffer concentrate to produce point-of-use buffer for the continuous capture operation)2 not only increases productivity, it also minimizes facility footprint, increases asset utilization, and reduces costs.

Figure 2. A two- to threefold increase in productivity is obtained when using continuous capture over single-column batch4

Sequential Batch Operation

The continuous operation of two interconnected downstream unit operations (e.g., flow-through and bind-elute polishing steps) is another way to increase productivity and streamline the downstream process. Two-fold increases in productivity for continuous, sequential batch operation, as compared to individual batch steps, has been reported.3  In addition, operating two connected polishing steps continuously eliminates the need for in-process hold and storage, and eliminates the time associated with changeover between individual unit operations—streamlining the process.

Operating sequential batch operations continuously on a two-column system, such as the EcoPrime Twin LPLC, combines two unit operations in one system, reducing the equipment footprint to maximize throughput per square foot of operating suite (Figure 3).

Figure 3. Data for two different loads shows a consistent twofold increase in productivity when using sequential over batch4 while maintaining a high yield

System Optimized for a Single-Use, Buffer Concentrate Environment

The EcoPrime Twin, with its onboard, highly precise (+/- 1.0% accuracy)2 buffer dilution option, provides:

• Use of buffer concentrates to save space
• Utilization of concentrated caustic for fully automated system CIP/sanitization
• Facilitates sequential processing, creating inline buffer for conditioning of feed between sequential two-column steps
• Significant space savings—two unit operations combined and use of concentrates further decreases tankage by 10-fold or more.

Single-Use Environment Enabled

A proprietary manifold by LEWA enables the unit to automatically and aseptically manage multiple buffers, WFI, and CIP from single-use bags/tubing sets. This arrangement allows for the use of the unit in a minimally classified area—significantly reducing costs associated with more highly classified cleanroom production environments. The adaptation to a commonly available single-use connector (DAC) simplifies the interface between the single-use and stainless-steel components on both the upstream and downstream sides of the unit. The ingenious design and automation assures the sanitization of all system-facing surfaces prior to opening the DAC, resulting in a fluid path that is never exposed to an open environment that is not sanitized.

De-Risk the Continuous Challenge and Gain Significant Flexibility and Productivity

Using this simple, patented design eases production implementation, validation, and maintenance. The new EcoPrime Twin Enhanced Platform delivers scalable, proven cost reductions (>50% Protein A and buffer reduction),4 significantly increases production flexibility, and provides two- to threefold productivity gains, as well as the ability to combine multiunit operations on a single skid in a single-use environment.

The simple design, along with options for multifunction enhancements, de-risks the investment both in validation ease and manufacturing asset utilization/flexibility. This is truly the Swiss Army knife for modern biomanufacturing facilities.

Wide Range in Commercial Production

Units with the functionalities mentioned above are now commercialized with flow ranges of up to 20 LPM—a platform enabling small to large production lot purifications on the same system.


Mary Jo Wojtusik, Ph.D. ([email protected]), is senior product manager, LEWA Bioprocess Technologies.

1. James Angelo, John Pagano, Thomas Müller-Späth, Kathleen Mihlbachler, Srinivas Chollangi, Xuankuo Xu, Sanchayita Ghose, and Zheng Jian Li, Scale-Up of Twin-Column Periodic Counter-Current Chromatography for MAb Purification, BioProcess Int. April 2018; 16(4).
2. Daniel Baur, Monica Angarita, Thomas Muller-Spath, Fabian Steinebach, and Massimo Morbidelli, Comparison of Batch and Continuous Multi-Column Protein A Capture Processes by Optimal Design, Biotechnol. J. 2016; 11(7): 920–931.
3. Mary Jo Wojtusik, Kurt Wilner, Low Pressure Liquid Chromatography Using EcoPrime LPLC with Enhanced Buffer In-Line Dilution, BioProcess J. 2016; 15(3): 14–19.
4. James Angelo, Srinivas Chollangi, John Pagano, Daniel Baur, Kathleen Mihlbachler, Thomas Muller-Spath, Xuankuo Xu, Massimo Morbidelli, and Sanchayita Ghose, Scale Up Design and Optimization for an Intensified Downstream Process Utilizing Multi-Column Operations, 255th ACS National Meeting, New Orleans, LA, March 2018.


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