Measuring and Quantifying Biomolecular Interactions are Critical in the Discovery and Selection of Biotherapeutic Lead Candidates
The high costs of discovery, development and production of drugs necessitate the constant need for improved process efficiencies.
Octet® Bio-Layer Interferometry (BLI) system implements a label-free analytical approach, which provides a fast and straightforward means to accurately measure the binding kinetics of biomolecular interactions, to screen library and to directly determine concentration of molecules of interest in their crude matrix. Its high throughput measurement capability make it powerful in every stage of biopharmaceutical development from early discovery to manufacturing.
Multiple examples highlights its advantages over HPLC and ELISA techniques in various segments of drug discovery and bioprocess development.
Unlike traditional surface plasmon resonance (SPR), BLI platform key features rely on “dip and read” plate format that do not use microfluidic systems to transfer samples to the biosensor surface. This provides greater flexibility in the types of samples used, without fluidic clogging concerns or the requirement of extensive sample preparation procedures (e.g cell lysates, aggregation-prone formulations…).
BLI Platform Key Features
Label-free analytical assay
Fluorescent labelling is not required, avoiding much time spent on sample preparation and reducing binding signal influence from the label itself.
Affinity dynamic range included between 10 pM and 1 mM
High flexibility in ligand capture method
In the presence of intrinsic tag or motif, a large choice of highly specific biosensor surface chemistries are available for the capture of ligand. When no tag is present, a coupling by direct immobilization can be performed.
Crude sample compatibility
Primary or secondary screening or quantification assay can be performed through the use of crude or complex samples (“analyte”) such as cell lysates, culture supernatants or solutions containing high refractive index components (e.g glycerol , detergent, DMSO…). Biosensors offers possibilities to capture ligand even from unpurified materials.
High flexibility of assay buffer conditions.
Quantitation Detection range down to 1-10 ng/mL
Real-time Kinetic Data Acquisition
Dynamic interactions are measured continuously during association and dissociation steps. The resulting traces are analyzed by exponential fit models to generate KD values but also highlight the difference between on and off rate constants for each intertaction. These results provide comprehensive data on specific interactions for decision-making.
High versatility in molecule orientation assignment
According to sought data, target or sample can be either captured on biosensors or used in solution. Tag availability, protein stability, orientation of interaction, multivalency effect, biosensors capture method, molecules size are primary concerns that guide capture orientation choice.
High-Throughput capacity
8 to 16 channels simultaneously in use enables to analyse binding kinetics of several dozen of compounds in one single experiment. The screening of several hundred of biomolecules can be performed in a single day, making Octet® BLI platform the highest-throughput option among label-free technologies
Dip and Read plate format
In absence of fluidic-based system, no risk of clogging when using crude or unpurified samples. Assay are performed in 96 or 384-wells plate instead of chip-trays as in typical SPR kinetics format and samples are not consumed or destroyed by analysis. These can even be recovered once the assay is completed.
Requires only nanomolar amount of samples making the approach suitable for challenging samples with limited availability.
High dynamic range of biomolecular interaction
Sensitivity range makes the Octet® BLI platform compatible to study various interaction, including protein-protein, protein-DNA/RNA, protein-small molecule, and even embedded target within particles. This versatility makes it applicable in many research and development contexts.
Dynamic sensitivity range from molecular weight of 1000 KDa (e.g virus) down to 150 Da.