An ever-increasing demand for biomolecule-based therapies is driving biopharmaceutical companies to accelerate process development activities to remain competitive in today’s industry. Within the downstream process development space, affinity chromatography is widely used for primary capture of monoclonal antibodies, Fc fusion proteins, etc. Optimization of this step requires determination of dynamic binding capacity (DBC) for target protein molecule and can be a lengthy process using traditional methods. KBI Biopharma has applied the use of variable pathlength technology for rapid generation of breakthrough curves and determination of DBC for capture resin selection. FlowVPE, a variable pathlength UV detector from C Technologies, is integrated with the chromatographic system for real time generation of breakthrough curves, reducing the time and cost associated with the development of the primary capture step. This technology helps accelerate the overall process development timeline. Case studies are included.

Antibody fragmentation and aggregation result in the formation of product-related impurities with the potential of immunogenicity, loss of biological activity and failure of internal and external regulatory standards. Conventional analytics rely on off-line HPLC, usually done well after the analyzed unit operation has ended and no process corrections can take place. Multi Angle Light Scattering (MALS) can provide the real time molecular weight (Mw) of a pure solute or the average Mw of a solute matrix if the total concentration of the species is also available in real time. Analytical experiments with low protein concentration can use conventional fixed path length UV sensors, however these become saturated at the higher concentrations of manufacturing processes. The FlowVPE System enables the real time measurement of protein content in a wide concentration range, which can be coupled to MALS for RT-MW measurements. Besides a more agile process development, RT-MW can enable adaptive processing decisions and inform on product deviations.

An evaluation of a beta version of the FlowVPX and Viper software was performed to determine it’s potential as an inline instrument for product concentration measurement during UF/DF operation, particularly high concentration UF/DF processes that are required for subcutaneous injection formulations (>150 g/L). The FlowVPX has significant potential for GMP and commercial biologics manufacturing unlike the first generation FlowVPE, which was designed as a lab scale instrument. Extensive testing was done using small and pilot scale UF/DF operations with several biologic drugs molecules from the portfolio. The FlowVPX instrument demonstrated very good comparability with other A280 based protein concentration measurements for all molecules tested (six). From a technical standpoint the FlowVPX is a GMP ready instrument, and further testing for instrument robustness and software development is ongoing at present to potentially use the instrument in the GMP space within the near future.

Protein concentration is one of the critical quality attributes of fill-finished drug products, as it ultimately defines the dosage a patient receives. Process analytical tools are key in monitoring this attribute and are used to control the manufacturing process accordingly. This presentation will give an overview of the journey from initial laboratory feasibility tests of the FlowVPE technology to implementation into an aseptic GMP facility for in-process control testing of biologics. In addition, the talk will highlight how the online technology sheds new light on the manufacturing process and increases product understanding. Building on the availability of real-time continuous protein concentration data, next steps in the development and implementation of a future automated human-free process execution will be shared.

Panel Discussion: Our panel with talk about the process to implementing and validating an in-line VPT system. They will discuss the planning methodology, the challenges they encountered and how they overcame them. They will also focus on the future of in-line testing, what is still missing? What are some exciting technologies on the horizon?