Creative Biolabs proudly provides you the Octet system from ForteBio for measuring antibody affinity. The Octet platform based on bio-layer interferometry (BLI) technology is a whole set of system including instruments, biosensors, reagents and assay kits to support the evaluation of biomolecular interactions in 96- or 384-well microplates. Octet system uses Dip-and-Read assay mode avoiding the need of microfluidics, and enables the real-time, label-free analysis of affinity as well as kinetics. Compared to the standard Biacore system, Octet has a higher throughput that provides greater potential in drug development applications. Additionally, Octet utilizes nm shift rather than response unit to define surface changes, which makes it much easier- and faster- in manipulation and cost-effective for antibody affinity measurement.
The principle of BLI technology is based on the optical interference pattern of white light reflected from two surfaces - a layer of immobilized protein and an internal reference layer. The binding between a ligand immobilized on the biosensor tip surface and an analyte in solution produces an increase in optical thickness at the biosensor tip, which results in a shift in the interference pattern measured in nanometers. The wavelength shift (Δλ) is a direct measure of the change in optical thickness of the biological layer, when this shift is measured over a period of time and its magnitude plotted as a function of time, a classic association/dissociation curve is obtained. This interaction is measured in real-time, providing the ability to monitor binding specificity, association rate and dissociation rate, and concentration with outstanding precision and accuracy.
Figure 1. The principle of BLI technology.
There are three biosensor-based assay orientations that can be used to explore antibody interaction (Figure 2): tandem blocking, premix blocking and classical sandwich. Compared to Biacore, Octet's dip-and-read assay allows longer analyte-binding steps and favors the rebinding of analyte to the ligand-coated sensors. Meanwhile, faster association time consumes less sample, which saves your precious proteins.
Figure 2. Outline of three biosensor-based assay orientations: (A) in tandem blocking; (B) premix blocking; and (C) classical sandwich.
Key Features of Octet system:
We can provide the following custom services using Octet system:
It is worth to mention that, if you need to compare a series of antibodies that target different regions with similar affinity constants, we are capable of perform epitope binning to evaluate each antibody separately. The antigen is captured onto the biosensor and the first antibodies are introduced to saturate the antigen, then the competing antibody is applied and measured with the kinetic curve getting determined to provide further characterization of the specificity. All of these could have significant effects for candidates' efficiency and pharmacokinetics evaluation.
Other optional Antibody Affinity Measurement Services:
Fig. 3 The binding affinity of bruceine A with p38α MAPK was determined by fortebio octet system assay and microscale thermophoresis. (Cai Lu, 2021)
Pancreatic cancer is one of the cancers with the worst prognosis, and people urgently need effective new chemotherapeutic drugs. Here, the researchers isolated a kind of quassinoid, bruceine A, from the dried fruit of Brucea javanica (L.) Merr., which showed strong anti-proliferation activity against pancreatic cancer both in vivo and in vitro. Proteomic analysis showed that p38α MAPK phosphorylation was related to the role of bruceine A in MIA PaCa-2 cells. The results of the Octet system and microscale thermophoresis assays showed that p38α MAPK had a high affinity for bruceine A. Molecular docking and molecular dynamics simulation experiments showed that bruceine A was widely bound to the P ring of p38α MAPK. The above results show that Brucein A can directly bind to p38α MAPK, which can be used to explore the role of p38α MAPK phosphorylation in the progression of pancreatic cancer and as a potential candidate for the treatment of pancreatic cancer.
The Octet system utilizes Bio-Layer Interferometry (BLI) technology to measure the affinity of antibodies to their antigens. This system monitors real-time binding interactions without the need for labeling, providing quantitative data on the strength of these interactions, which is crucial for determining the effectiveness and specificity of antibodies in therapeutic applications.
Measuring antibody affinity is critical in therapeutic development to ensure high specificity and efficacy of antibody-based drugs. High affinity antibodies are more likely to bind to their specific target with greater strength and stability, which is essential for achieving desired therapeutic effects and minimizing off-target interactions.
Unlike ELISA, which typically measures the overall binding capacity and requires labeling, the Octet system measures binding kinetics in real-time without the need for labels. This allows for a more direct assessment of the binding strength (affinity) and kinetics, including on-rate (association) and off-rate (dissociation), providing a more comprehensive understanding of how an antibody interacts with its antigen.
Octet system can measure antibody affinity directly in crude samples such as cell culture supernatants, serum, or plasma. This capability significantly streamlines the development process by allowing the measurement of antibody interactions without the need for extensive sample purification, thus saving time and resources.
The Octet system can analyze a wide range of antibodies, including monoclonal, polyclonal, and recombinant antibodies. It is versatile enough to handle various formats such as full-length IgG, Fab fragments, and single-chain variable fragments (scFvs), making it suitable for diverse research and therapeutic development needs.
The Octet system is highly sensitive and capable of detecting even low-affinity interactions. Its technology allows for the measurement of a wide range of affinities, from very high to very low dissociation constants (KD), enabling comprehensive profiling of antibody interactions across different therapeutic and research applications.
Real-time measurement in the Octet system offers several benefits for antibody development, including the ability to observe the kinetics of antibody-antigen interactions as they occur. This provides immediate feedback on the binding characteristics, such as association and dissociation rates, facilitating rapid optimization of antibodies for improved performance in therapeutic applications.
Octet system is well-suited for conducting comparative studies of different antibody batches. By measuring and comparing the affinity and kinetic profiles, researchers can ensure the consistency and quality of antibody production, which is critical for maintaining efficacy and safety in therapeutic applications.
Use the resources in our library to help you understand your options and make critical decisions for your study.
All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.
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