Method Development and Qualification Considerations for Octet-Based Potency Assays


The pressure to improve speed to market directly impacts the analytical assay development timeline. The current industry standard from target to phase I clinical trials is 18 months, with companies such as KBI Biopharma targeting 8 months.  Potency assays are often time consuming to develop and require significant investment to demonstrate reproducibility. With the advent of BiAb therapy, dual binding potency assays have also become critical to successful filing. Developing a dual binding assay that demonstrates binding to two separate antigens is not only technically challenging, it requires even more time to develop. KBI Biopharma has rapidly developed a variety of Octet® based potency assays and successfully qualified the methods. Throughout development and qualification, there are many important factors to consider such as method type, loading factors, accuracy, precision, specificity, range, linearity, and parallelism. The Octet® platform has the ability to assess each of these parameters rapidly, supporting accelerated timelines.

Automated Optimization of IgG Production in CHO Cells

2017-Aug-29 to

Antibodies are frequently produced from a cell line that has been screened to ensure the protein is expressed with the desired post-translational modifications, target specificity and affinity, and at relatively high levels. The identified cell line can be further developed to achieve higher titers and to increase cell growth and/or protein production through the optimization of the cell culture media. An ideal approach to media optimization is through the use of a factorial design of experiment (DOE), where a variety of media components are tested at different concentrations and in combination with one another. The challenge though, is that these factorial experiments rapidly increase the number of conditions that require testing. It is desirable therefore to use instrumentation that can minimize the time required for sample preparation and quantification during the optimization process.

Common ways of quantifying the production of IgG or other proteins are either labor-intensive (e.g. ELISAs) or prohibitively slow (e.g. HPLC), particularly at the throughput required for DOE studies. A rapid alternative for quantification and kinetics analysis of biologics is the high-throughput Octet HTX platform that utilizes Bio-Layer Interferometry to detect real-time binding of molecules.  Here we show how coupling this high-throughput analysis with the Biomek FXP Automated Workstation enabled the DOE optimization of IgG expression in a CHO cell line. The Biomek FXP Workstation was used to prepare 96 media combinations and plate cells in replicate conditions, generate sample and reagent plates for the Octet HTX system, and initiate XTT assays to better understand the effect of media components on CHO cell growth.

Comparative Fragment Screening Approaches for ClpP Activators using the Pioneer FE SPR System

2017-May-25, 11:00 AM to

Fragment-based drug discovery requires higher throughput to process large numbers of fragments within a short amount of time and high sensitivity to reliably detect the weak affinity interactions of low molecular weight analytes. The Pall ForteBio Pioneer FE system lets you screen for identification of fragment candidates on large numbers of sample and obtain reliable affinity (KD) and kinetic (ka, kd) data directly from the primary screens.

This webinar will provide an overview of the Pioneer platform technology and its application in a real-life discovery workflow.

Dr. Aman Singh will present the results of a 5000 fragment screening and hit triaging effort to find alternative ClpP binders using the Pioneer FE system. Caseinolytic Protease P (ClpP) represents an important emerging target for the treatment of chronic bacterial infections. Acyldepsipeptides (ADEPs) specifically target the ClpP allosteric activation domain resulting in deregulated hyper-activation of ClpP, leading to self-digestion of pathogenic bacteria. A retrospective analysis of success of Surface Plasmon Resonance (SPR), Thermal Shift Assay (TSA), and Fluorescence Polarization (FP) assays against this complex protein-protein interaction site target will also be presented.

One Step Ahead for Fragment ScreeningGEN Webinar - Cell-Based Label-Free Monitoring of Signal Transduction


10 AM ET, 7 AM PT
Dennis Verzijl, Ph.D., Genmab
Craig Tin, Director, Regional Marketing, Pall ForteBio

Label-free technologies provide critical information for the drug discovery process, from the screening of specific drug target binding interactions, to measuring the affinity of that binding. When used with purified proteins or membrane fractions, these assays are limited in providing rich insight into the complex milieu of cellular response signaling molecules. Bio-Layer Interferometry (BLI) has recently been demonstrated to monitor binding interactions in real time with live cells, creating a novel cell-based BLI that has the potential to increase the versatility of label-free assays in screening labs for finding hit that rely on more complex signal transduction.

This webinar will describe the development of a cell-based BLI assay on the Pall-ForteBio Octet HTX system and explore how high-throughput label-free assays can streamline the drug discovery process.

Predicting Human IgG Half-Life with an In Vitro Assay


Colby Souders, Ph.D.
Kanyos Bio
10 AM EST New York/3 PM BST London

Immunoglobulin G (IgG) has an unusually long serum half-life in comparison to proteins of similar size due to IgG’s ability to bind the neonatal Fc receptor (FcRn) in a pH-dependent manner. FcRn binding properties can vary among IgGs, resulting in altered in vivo half-lives, making it beneficial to accurately predict the FcRn binding properties of therapeutic IgG monoclonal antibodies (mAbs). This webinar will describe the development of an in vitro model capable of predicting the in vivo half-life of human IgG. Using a high-throughput Bio-Layer Interferometry (BLI) Octet® platform, the human FcRn association rate at acidic pH and subsequent dissociation rate at physiological pH was determined for 5 human IgG1 mAbs. Comparing the combined FcRn association and dissociation rates to the Phase 1 clinical study half-lives of the mAbs resulted in a strong correlation. The correlation was also verified in vivo using mice transgenic for human FcRn.

The model was used to characterize various factors that may influence the FcRn-mAb binding, including mAb variable region sequence differences and constant region glycosylation patterns.

Biomolecular Binding Kinetics and Affinity Determination Using Bio-Layer Interferometry

2016-Mar-17 to 2016-Mar-18

The Octet system from Pall ForteBio offers a rapid, versatile, and sensitive label-free solution for measuring kinetics of biomolecular interactions. One of the most challenging aspects of such measurements is the reliable analysis and interpretation of kinetic data. In this webinar, you'll learn:

  • The basics on binding kinetics experiments
  • Step-by-step data interpretation
  • How to analyze binding curves
  • Best practices for generating graphs for publications


Webinar: Fast and Flexible Analysis of Fc-Gamma Receptor Binding Interactions on the Octet PlatformWebinar: Cross-competition or Epitope Binning Assays on the Octet HTX SystemWebinar: Advances in High-Throughput Protein Analysis: The New Octet HTX SystemWebinar: Achieving More with Less - Unlocking the Versatility of Your Octet System with Different BiosensorsImproved HIS-tagged Protein Detection with ForteBio's NEW HIS2 Biosensor


Presenter: Darick Dayne, Ph.D., Senior Product Manager, Pall ForteBio

Best Practices for Large Molecule Kinetic Assays on the Octet System


Label-free analytical technologies such as the Octet® system provide a powerful means to obtain information about biomolecular binding interactions and kinetics, key components of a drug-target interaction. Presenter: Renee Tobias, Applications Scientist, ForteBio - A Division of Pall Life Sciences

Webinar: Getting the best data from your BLItz label-free systemUsing the BLItz personal assay system to overcome bioprocess development bottlenecks


Presenter: Hendrik Wünsche Ph.D., Applications Scientist, ForteBio - A Division of Pall Life Sciences The BLItz system is a simple-to-use tool for protein analysis that works with only a drop of sample. This webinar will provide a brief overview of the BLItz system and the capabilities it enables in a bioprocessing environment.

Kinetics on the Octet Systems: What Lies Beneath the Curves


Presenter: Phil Buckle, Application Manager, Europe, ForteBio - A Division of Pall Life Sciences In this webinar, we will discuss kinetics analysis on the Octet and BLItz Systems.

Molecular Binding Kinetics: What Lies Beneath the Curves


Presenter: Phil Buckle, Application Manager, Europe, ForteBio - A Division of Pall Life Sciences Molecular interactions play a myriad of important roles in drug discovery and development. In this webinar, we will discuss kinetics analysis on the Octet and BLItz Systems.

Octet Platform: Versatile Instruments in Downstream Processing


Presenter: Phil Buckle, Application Manager, Europe, ForteBio - A Division of Pall Life Sciences The large number of biotherapeutic molecules in development has resulted in dramatically increased sample workloads for analytical testing laboratories. To address this bottleneck, many biopharmaceutical companies are adopting a new generation of rapid, high-sensitivity analytical techniques for process characterization and monitoring.

Webinar - Detecting E. coli O157:H7 Using KPL BacTrace Antibodies and the Octet RED96


Presenter: Brian Bullard, Ph.D., Research Leader - Bacterial Detection, KPL, Inc. Early detection of foodborne pathogens is the primary means of preventing contaminated food from reaching the consumer. Traditional immunoassays, such as Western blot and ELISA, are specific but time-consuming due to required enrichment processes. Newer detection methods, such as multiplex PCR, improve on the specificity of traditional methods but require 1-2 hours per assay in addition to several hours of pathogen growth prior to testing.

Webinar - Epitope Binning Strategies in Therapeutic Antibody Development

2012-Sep-25, 8:00 AM

Presenter: David O. Apiyo, Ph.D., Field Applications During therapeutic antibody development, panels of monoclonal antibodies are routinely screened for their epitope binding regions. Antibodies that bind to the same or overlapping regions of an antigen are categorized into epitope groups or "bins." Depending on the design of the assay, results of epitope binning assays can be difficult to interpret. In this webinar, we will discuss techniques for label-free epitope binning studies and methods currently in use. The strengths of different assay formats will be considered. Approaches to antibody pair selection will also be discussed.

Webinar - How to Overcome Bioprocess Development Bottlenecks Using ForteBio's BLItz System

2012-Sep-12, 8:00 AM

Presenter: Renee Tobias, Applications Scientist BLItz is a highly accessible, simple to use protein analysis tool that works with just a drop of sample. It uses a Drop. Read. Done! format to characterize proteins instantly. Complex protein samples are analyzed to: Report the presence or absence of a specific protein in just a few seconds. Accurately measure the concentration of proteins with excellent sensitivity and specificity. Monitor the binding of biomolecules in real-time to report binding rate and affinity constants. Provide more information about each step involved in ELISA assays to help develop better ELISAs faster and easier.

WEBINAR: ForteBio's new Anti-Human Fab-CH1 Biosensor, technical tips on usage and applications


Presenter: Darick Dayne, Ph.D., Product Manager, ForteBio - A Division of Pall Life Sciences ForteBio launches the new Anti-Human Fab-CH1 Biosensor for easy quantitation and kinetic measurements of human Fab, F(ab')2, and IgG. Its unique set of specificity makes it a versatile tool for a wide range of applications from bioprocessing to product QC.

WEBINAR: CaptureSelect affinity ligands on ForteBio's Octet platform for antibody characterization and quantitation


Presenter:Pim Hermans, BAC BV, Director of Ligand Discovery Since more and more antibody based therapeutics are lacking a regular Protein A binding site it becomes a challenge to find a Protein A equivalent for primary capture of antibody formats. For this reason BAC has developed a range of CaptureSelect affinity ligands directed against a unique panel of antibody sub-domains (e.g. present on Fab) to facilitate affinity purification and detection of virtually any antibody based format.

WEBINAR: Protein Concentration Measurements in Crude Samples in a Dip and Read Format


Presenter: Philip Buckle, ForteBio Applications Manager, Europe Abstract: Learn how you can use the Octet and BLItz platforms to rapidly develop and run concentration measurements of proteins in crude solutions. Bio-layer interferometry technology and experimental design strategies will be discussed, with examples including recombinant protein, IgG and Fab quantification

Webinar: Protein Concentration and Kinetic Characterization Measurements of Crude Samples in Dip and Read Format

2012-Jun-18, 11:00 AM

Presenter:Yixin Lin, Ph.D, Field Applications Scientist Learn how you can use the Octet or BLItz platforms to rapidly determine the concentration of antibodies and proteins in crude solutions and investigate their binding kinetics in a label-free format. Bio-layer interferometry technology and experimental design criteria will be discussed. This webinar will provide examples of IgG and HIS-tagged protein characterization as well as review select publications citing use of the Octet platform for crude sample analysis.

User Video Presentations

View Octet User video presentations, user presented webinars and ForteBio's applications scientists presented webinars.

Fast, Label-free Assay for Quantitation of a Humanized Antibody Therapeutic
Presented by Mark Dysinger, Senior Scientist, Pfizer Global R&D
BLI is an established technology for evaluating the kinetic properties of ligand binding reagents and protein therapeutics, with practical utility to quantify biotherapeutics, for example in toxicokinetic (TK) studies. An Octet assay for the quantitation of a humanized antibody therapeutic with validated ELISA reagents is discussed along with quantitative use of BLI in both non-GLP and GLP environments.

(Click to view)

Fully Automated CHO Host Cell Protein Quantitation in Monoclonal Antibody Therapeutics
Presented by Dan Schuessler, Analytical Scientist, GlaxoSmithKline
The quantitation of CHO Host Cell Proteins in monoclonal antibody therapeutics using a laborious sandwich ELISA assay takes approximately 5.5 hours to test one 96-well assay plate. With minimal effort, an Octet CHO HCP quantitation assay was developed that can be completed in approximately 2 hours, allowing up to three 96 well assay plates in a single run, hands-free after the initial setup.

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