Applications


Applications

The applications of the BLI-technology and the Forte-bio Octet instruments are many. Choose your grey topic of interest on the image below.

BLI - applications
Applications
Kinetics Glycosylation ADCC Pharmaconetics Potency

Kinetics

Use BLI for rapid kinetic measurements. Determine de ka, kd and KD in one run. Because BLI is not sensitive to the liquid matrix containing your sample, screening of un-purified samples is simple and easy by just dipping the sensor in. In addition, construct affinity isotherm plots to rank the samples and determine which are best suited for your application.

Characterise your antibodies with epitope binning to select the ones with different epitopes, perform isotyping and select antibody pairs which can be used in ELISA’s.

Look for more information at Kinetic Characterization.

Antigen-Antibody

Use BLI to characterize antibodies from hybridoma supernatants, phage lysates or purified samples in real time, while eliminating any sample preparation beyond an optional dilution step. Octet instruments can read from 2 to 96 samples simultaneously in automated format using a standard microplate for rapid determination of binding affinity constants (KD), association rates (ka or on-rate) and dissociation rates (kd or off-rate) for antibody-antigen binding. In addition to complete kinetic characterization, equilibrium assays can be used to determine KD, or simple binding assays performed to rapidly evaluate relative affinity or screen large numbers of samples. BLI biosensors are compatible with a wide range of sample types and buffers, including cell extracts, serum, and media. The standard microplate format combined with disposable Dip and Read biosensor technology enables automated, highly parallel processing on the Octet system in sample volumes as low as 40 µL.

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Glycosylation

Drug product glycosylation is a critical quality attribute (CQA) due to its potential impact on pharmacokinetics properties and stability of the product. ForteBio’s Sialic Acid (GlyS) Kit enables high-throughput relative screening of sialic acid content in crude and purified samples. There is no need for sample purification or glycan digestion steps. 1000 clones can be screened in just under 10 hours on the Octet HTX system.

A combination of the Sialic Acid (GlyS) Kit and Protein A biosensors, or any of the ForteBio quantitation biosensors can be used to perform titer and sialic acid content screening on the same samples using Octet systems. Octet Data Analysis HT software allows titer data to be combined with sialic acid content data. The ability to view and choose from desired titer and sialylation levels at the same time provides more in-depth knowledge that facilitates more informed decisions.

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ADCC

Antibody-mediated cell-dependent cytotoxicity (ADCC) (1) is the major way of action of therapeutic monoclonal antibodies. The three classes of Fc gamma receptors (FcγRs) are membrane glycoproteins with affinity for the Fc region of immunoglobulin G (IgG). Induction of ADCC by an antibody depends on its binding affinity to both the target and the FcγR. Monitoring the maturation of the antibodies is of great importance since specificity and high affinity is beneficial for safety and efficacy of therapeutic antibodies.

BLI makes it possible to check the interaction between the FcγR – IgG – Target in one experiment. This gives valuable information on the individual kinetics and the overall kinetics of the system.

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Pharmacokinetics

High detection sensitivity, a wide dynamic range, and streamlined workflows that shave hours or even days from standard processes make the Octet system a compelling alternative to ELISA for pharmacokinetic and pharmacodynamic studies. Anti-drug immune responses can be precisely characterized in serum samples, providing a detailed profile of antibody isotype, specificity, and affinity.

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Potency

The Octet platform offers a fast, accurate, and robust solution for measuring potency of a drug molecule. The speed of the Octet systems allow for rapid Design of Experiment results which can aid in development, optimization, and potential validation practices. The system is ideal for characterizing biotherapeutic activity and monitoring potency and stability.

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Binding kinetics assays and specifically affinity constant (KD) analysis are increasingly being used for biological products lot release. Regulatory requirements necessitate that such products be QC tested using methods that are robust and have been appropriately developed, qualified and validated under GMP conditions. Octet platforms come with a complete GxP package that include instrument installation and qualification (IQOQ) kits, performance qualification (PQ) kits and 21 CFR Part 11-compliant software. In addition, we provide users with software and biosensor validation support that facilitates compliance with GMP requirements.

Rely on rapid development, optimization and validation of assay methods for various applications under GLP and for QC laboratories.

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References

(1)Nimmerjahn, F. and J. V. Ravetch Fc[gamma] receptors as regulators of immune responses. Nat Rev Immunol 8: 34-47; (2008). Goto reference