Accelerating the Development and Manufacture of Therapeutics
The discovery, development, and commercial manufacturing of viral vectors, vaccines and other biotherapeutics can be a costly and lengthy endeavor. To shorten development timelines and bring novel therapies to market faster, rapid analytical tools are needed to support characterization, testing, and optimization activities necessary to achieve robust and cost-effective manufacturing. Sartorius offers the Octet® BLI platform based on biolayer interferometry (BLI) technology providing an alternative to conventional analytical techniques to support a wide range of bioprocessing-based applications. BLI technology can overcome the limitations in throughput, performance, and cost associated with traditional analytical platforms like UV spectroscopy, enzyme-linked immunosorbent assays (ELISAs), and high-performance liquid chromatography (HPLC).
It is an advanced, label-free, fluidics-free optical biosensor that can quantify biomolecular interactions by measuring changes in the interference pattern of white light reflected between a biosensor tip and an internal reference layer. The plate-based, fluidic-free design is equipped with biosensors coated with immobilized ligand that are dipped directly into the sample kept in a 96- or 384-well plate format. The specific binding of an analyte in the sample to the immobilized ligand on the sensor surface produces a wavelength shift in the interference pattern that can be detected in real-time. This enables rapid analysis of protein interactions, steady-state affinity, quantification, binding kinetics and epitope sites in complex matrices and in crude samples, without the need for purification or complex sample preparation. This unique characteristic makes BLI ideal for in-process sample testing, greatly increasing analytical capabilities during production while also reducing data turnaround time to provide significant time and cost savings over traditional analytical assays.
The Octet® BLI platform and assays are well-suited to execute real-time, high throughput, automated analysis across process development and manufacturing operations to accelerate workflows and shorten timelines. A recent report published by BioProcess International entitled Accelerating the Development and Manufacture of Therapeutics Using the Octet® Platform provides a detailed explanation of BLI principles and explores the benefits in upstream and downstream bioprocessing for the development of viral vectors and vaccines. Here, we summarize several key findings from the report.
Bioprocess Development Applications
Optimization of upstream and downstream manufacturing unit operations during process development is essential for successful and cost-effective production of biotherapeutics. In upstream development, the Octet® BLI platform can be used to facilitate a broad spectrum of activities from hybridoma clone screening and selection, cell-line development, protein titer determination, to assessing the influence of process changes on product critical quality attributes (CQAs) during optimization. Additionally, researchers can leverage preconfigured reagents and protocols for rapid protein quantification (protein products, host-cell proteins, residual protein A) to determine the impact of multiple process variables to define optimal downstream purification conditions.
|Upstream Applications||Downstream Applications|
|Concentration Measurement||Dynamic Binding Capacity|
|Kinetic Rate||Detection of Residual Contaminants|
|Affinity Analysis||Formulation Development|
|Titer Determination/Clone Selection||Quality Control (i.e., activity, potency)|
|Cell Line Development||Stability Testing|
Viral Vector Bioprocess Applications
During viral vector development, characterization, and quantification of adeno-associated virus (AAV) particle characteristics such as capsid titer, empty-full capsid ratio and genome titer employs several analytical assays such as ELISA, analytical ultracentrifugation (AUC) and droplet digital polymerase chain reaction (ddPCR). These methods are effective, yet they are too time-consuming and impractical for at-line measurements during production. A BLI-based AAV assay, which uses a biotinylated capture molecule, has the versatility and speed to quantitate different AAV serotypes and test hundreds of process samples with minimal matrix interference to overcome some of these challenges. In direct comparison with ELISA quantification, it reduced time to results by 80%, operator hands-on time by 50%, and sample preparation time by 15‑25% to enable rapid, at-line measurements during AAV production.
Vaccine Bioprocess Applications
Reducing development time and increasing the speed to market for novel vaccines has never been more critical. Label-free biomolecular interaction analysis accelerating vaccine development workflows is highlighted in several examples including SARS-CoV-2, influenza, and dengue fever across key milestones, from lead candidate selection, characterization of epitope diversity and coverage, to quantifying titer, potency, and executing stability measurements.
The Octet® BLI platform provides users with a highly versatile and easy-to-use analytical tool to accelerate the development and manufacturing of biotherapeutics in support of Quality by Design (QbD) strategies that drive processes efficiencies and cost-savings. The ability to monitor kinetics, affinity, and antibody/protein quantitation in crude samples, in real time at high throughput decreases experimental time and enables rapid process optimization while ensuring product performance. The addition of the Octet to the process development analytics toolkit can help create sustainable, scalable and robust manufacturing bioprocesses for viral vectors, vaccines and other lifesaving biotherapeutics.
To download the full report, please see Accelerating the Development and Manufacture of Therapeutics Using the Octet Platform.
For more information on label-free protein-protein and protein-small molecule interaction analysis, visit www.sartorius.com/octet