Overview for Biologics GxP Applications
Regulatory requirements necessitate that biologics drug products be QC tested using methods that are robust and have been appropriately developed, qualified, and validated under GMP conditions. Among these assays, titer measurement for potency assessment and ligand binding kinetics assays are increasingly being used for biological products' lot release. Drug developers are constantly in search of analytical technologies with robust features that are easy to use and adaptable in QC and manufacturing.
Fc receptor binding assays on the Octet system
The safety and efficacy of a therapeutic monoclonal antibody can be greatly impacted by its ability to bind to both the target and to the FcγR. Antibodies are often selected based on their binding properties to FcγRs. They are sometimes engineered to achieve desired FcγRs binding properties. As a result, the assessment of binding affinities of these antibodies to FcγRs is an integral part of biotherapeutic development processes.
- Octet systems offer high throughput and sensitive methods for Fc receptor binding analysis
- A variety of biosensor surfaces are available and allow for flexibility and rapid optimization of assay format
Ligand binding potency assays on the Octet platform
Reproducible and accurate relative ligand binding potency methods can be developed using analytical technologies. These methods can be used to asses lot-to-lot variability by monitoring either the kinetics of binding or by comparing affinity constants. They can also be used to determine the stability of the different lots of the drug, hence the method has to indicate stability. Due to their ease of use and fast time to results, real-time label-free analytical techniques are increasingly becoming attractive for the development of methods for assessing the potency of drug candidates.
- Readily accommodates the use of DOEs to rapidly develop robust potency measurement methods
- Develops relative affinity constant or response signals comparability studies for potency measurements
Titer determination on the Octet platform
Titer and protein concentration determination is a critical process in the development of biologics drug molecules. The active protein concentration can be used to determine the potency of the drug molecule. While ELISA and especially HPLC are commonplace techniques for titer and protein concentration determination, techniques that are more robust to cell culture and media are especially desirable as they can be easily adopted in both upstream and downstream processes during the development of the drug molecule.
- A full plate (96 samples) of IgG titer can be analyzed in as little as two minutes
- Sample plate format allows for the use of crude and non-purified samples
- Automation capable Octet HTX and Red 384 allow for walkaway high throughput analysis
Resources of GxP Applications
Application Note Enhancing Efficiency and Economics in Process Development and Manufacturing of Biotherapeutics
Application Note Validated Quantitation and Activity Assay of Antibody Fragment Molecule (Fab) for Process Development and Quality Control
The group was able to develop a working Fab activity assay on the Octet RED system in less than a week. Relative to the overnight incubation and four-hour assay time of their ELISA protocol, the Octet assay provided an analysis time of only one hour per 96-well microplate, including sample preparation time. This Octet assay was used to monitor Fab activity for all process development studies.
Application Note Analysis of Fc-gamma Receptor-IgG Interactions on the Octet Platform
Fc gamma receptors (FcγRs) are membrane glycoproteins with affinity for the Fc region of immunoglobulin G (IgG). FcγRs expressed on the surface of immune effector cells play a key role in initiating Fc effector functions such as antibody-mediated cell-dependent cytotoxicity (ADCC)1, which is a major mechanism of action of therapeutic monoclonal antibodiesRead more
Application Note Analysis of FcRn-Antibody Interactions on the Octet Platform
The Fc region of human IgG contributes to a number of beneficial biological and pharmacological characteristics of therapeutic antibodies.Read more
Application Note Octet Potency Assay: Development, Qualification and Validation Strategies
Kinetic analysis of biomolecular interactions is critical during drug discovery and development. The affinity of an interaction directly affects the dose required for a biopharmaceutical to be effective.Read more
Flyer Quality control of biotherapeutics using Octet systems
Bio-Layer Interferometry (BLI) has been rapidly adopted as an important analytical tool in laboratories that work with biological molecules, either as drug products, vaccines, or diagnostic reagents.Read more
White Paper Octet platform: a complete solution for lot release and in-process testing of biologics in GxP laboratories
Bio-Layer Interferometry (BLI) is an optical analytical technique that analyzes the interference pattern of white light reflected from two surfaces: a layer of immobilized protein on the biosensor tip and an internal reference layer (Figure 1A). Any change in the number of molecules bound to the biosensor tip causes a shift in the interference pattern that can be measured in real time (Figure 1A and 1B). The binding between a ligand immobilized on the biosensor surface and an analyte in solution produces an increase in optical thickness measured as a wavelength shift, Δ λ (Figure 1C).Read more