Free eBook: Label-free detection: technologies, key considerations, and applications

Real-time analysis of biomolecular interactions close to native biological conformations

FortéBio by Molecular Devices (Germany) GmbH

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Kinetics characterization using label-free detection can readily replace label-dependent methods such as ELISA or fluorometric methods. Real-time biosensor assays allow to gauge progression of the assay and also dissect binding affinities and mechanism of action with information from association (ka) and dissociation (kd) kinetics which ELISA does not provide.

Kinetic & Affinity Characterization - While ELISA has been the dominant technique in affinity characterization of biomolecules, real-time label free technologies have become increasingly popular due to their ability to provide more detailed information in the kinetics of binding of biomolecules and faster more sensitive quantitation results.

Cell Line Development – Biosensors are used to develop stable cell lines which are widely used in a number of important applications including biologics (e.g. recombinant protein and monoclonal antibody) production, drug screening, and gene functional studies.

Bioprocessing - Titer & Impurity Testing - From upstream processes such as cell line development to downstream processes like purification columns dynamic binding capacity assessment, product contaminant testing, titer, etc, label-free analytical techniques that enhance efficiencies while maintain-ing quality are critical to the development of biopharmaceutical drugs.

Interactions between biomolecules serve as key triggers for many biological processes and, therefore, provide perfect targets for drug discoveries. Biological binding interactions are a dynamic process driven by changes to the environment. Therefore, techniques used to characterize these interactions needs to mirror the level of biological complexity in order to fully understand these systems.

Commonly used label-free binding analysis platforms include Bio-Layer Interferometry (BLI), Surface Plasmon Resonance (SPR), Isothermal Titration Calorimetry (ITC) and Microscale Thermophoresis (MST). In this e-book, the four technologies are compared in terms of their capabilities and workflows to provide guidance for choosing the most suitable platform based on assay needs and application.

Key attributes such as throughput, sample capacity, unattended runtimes, affinity ranges, sensitivity, type of sample matrix and ease of use are important considerations when choosing a biosensor platform to suit your application and workflow needs.

Both the Octet BLI and Pioneer SPR platforms are capable of characterizing a wide range of biomolecular interactions. Pioneer SPR systems are ideal for small molecule and fragment screening workflows (> 70 Da) due to their exquisite sensitivity and next-generation injection capabilities. Octet BLI platforms can be used for analytes > 150 Da and applications involving larger molecular weight analytes such as viruses, nanoparticles, liposomes, and cells.

Octet systems are microfluidics-free providing an essential tool for routine analysis of crude lysates or complex matrices. Crude sample compatibility is useful especially in epitope binning, quantitation, off rate ranking or large screening assays where sample purification is neither required or feasible for efficient workflows.

Facts, background information, dossiers
  • real-time analyses
  • label-free analysis
  • bio-layer interferometry
  • surface plasmon resonance
  • isothermal titratio…
  • MicroScale Thermophoresis
  • interferometry
  • calorimetry
  • thermophoresis
More about FortéBio
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