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Confident antibody selection with Biacore™ 4000

Antibodies and their derivatives are increasingly important as biotherapeutics, precision diagnostics and essential tools for biological research. Technological advances now enable the production of large numbers of candidate antibodies. Techniques to screen and characterize large numbers of antibodies and provide confident selection of the best candidates for development are therefore essential. Biacore 4000 is designed to meet these challenging demands:

 • High quality, real-time data based on biologically relevant  binding properties (selectivity, affinity, kinetics)
 • Rapid analysis direct from crude samples (e.g. hybridoma supernatants, cell lysates) to avoid critical bottlenecks
   in the workflow
 • Low consumption of  potentially limiting samples and target antigens
 • Hardware and software designed to provide the throughput required for large-scale antibody analysis

As described below, GE Healthcare also provides an add-on software package with dedicated tools to enable customers to fully exploit the capabilities of the system for antibody analysis applications.

Biacore 4000 Antibody Extension Package

A powerful solution for large-scale antibody analyses
This software package enables Biacore 4000 users to get the best out of their system for antibody analysis applications,  saving time and reagents in biotherapeutic development. Dedicated software tools for high throughput kinetic studies and epitope mapping enable rapid identification of promising candidates:

• Obtain decision-critical kinetic information early during screening using
   2-over-2 kinetics 
• Save days during pairwise epitope mapping
• Address a broader range of antibody-antigen interactions using
  additional kinetic fitting models
• Conveniently  export data to analysis software such as Spotfire™ or
   Excel™ for customized evaluation and simplified workflow integration.

The main  technical features and associated application benefits  provided by the software package are described in more detail below.

• Go directly to: 2-over-2 kinetics>>

• Go directly to: Kinetic models>>

• Go directly to: Epitope mapping>>

• Go directly to: Literature download>>

• Go directly to: Data export>>
Additional spot-addressing functionality to meet the needs of antibody characterization assays
Underlying two key applications provided by the software package, additional spot-addressing functionality makes full use of the innovative flow system in Biacore 4000, enabling a valuable new approach to kinetic analysis, streamlined, reagent-efficient epitope mapping, more efficient assay setup and saving time and reagents during antibody screening and characterization.

Fast and efficient antibody characterization: 2-over-2 kinetics
Utilizing the additional spot-addressing functionality provided in the software package, the 2-over-2 approach to kinetic analysis was developed to further optimize efficiency in information-rich antibody screening and characterization. As shown opposite, the method captures two densities of two different antibodies in each flow cell, which are then analyzed using just two concentrations of antigen. Advantages of this approach include:

• Capture-based  approach minimizes assay development time
• Twice as fast as traditional multi-cycle kinetic assays, up to
  200 antibodies/24 hours
• Higher throughput for kinetic characterization without compromising on
  data quality
   - increases productivity e.g. during phage display workflows
• Kinetic analysis of antobodies directly from small volumes  of crude
  samples (<200 µl)
   - Confident selection of clones during early screening overcomes
      bottlenecks e.g. during early phases of hybridoma-based workflows

Asssay setup for 2-over-2 kinetics
2-over-2 kinetic evaluation

Streamlined & reagent-efficient epitope mapping
Dedicated tools for large-scale epitope mapping assays provide support for setting up, running, and evaluating pairwise epitope mapping experiments. Predefined methods for assay setup and evaluation are supplied, which can be used directly or modified as required, providing flexibility in assay design. Utilizing the additional spot-addressing functionality available, the epitope mapping tools enable highly efficient assays, saving significantly on time and reagents.

• Automatic generation of the most efficient run setup optimized for run
   time & reagent consumption
   - Based on one of two main capture formats (see opposite)
   - Minimizes hands-on time and potential errors
• Simple six-step workflow guides the user through evaluation of data
   - Binders identified using threshold levels and off-rates estimated 
   - Results displayed simply and clearly in a color-coded matrix table 

Setup alternatives for epitope mapping
Display of epitope mapping results

Extended export functionality for flexible data analysis
An extensive export functionality streamlines workflow integration and allows customized data analysis. Detailed information  is exported to visualization software such as Spotfire™ and Excel™, including run and sample information, curve markers, and evaluation results. Baseline adjusted sensorgram data can also be conveniently exported and displayed in Spotfire.

• Enables combination and summarization of data for more confident, faster decision making
• Saves time and resources
• Simplifies overview of multiple projects

Additional kinetic fitting models for broader applicability
The provision of additional fitting models broadens the application space for the characterization of antibody-antigen interactions. These may vary in their binding mechanisms and hence their requirements for kinetic fitting. Biacore 4000 Antibody Extension Package includes the following models:

• 1:1 dissociation: fits the dissociation phase when the analyte binds to one interaction partner attached to the surface
• Heterogeneous ligand: when the analyte binds independently to heterogeneous interaction partners or binding sites on
   the surface
• Bivalent analyte: used for analytes with two binding sites

Download Product & Application Literature

To download the pdf's below, please enter your login details. If you do not have a Biacore website account yet, you can sign up here.

Biacore 4000 Antibody Extension Package Data File (842 kB)
Application Note:
Resolving a bottleneck in screening and characterization of recombinant antibody fragments using Biacore 4000-AA.pdf (2 MB)