High Throughput SPR for Fragment Based Drug Discovery
- High Throughput Fragment Based Drug Discovery - The Pioneer FE system incorporates a high throughput mode which can determine the kinetics and affinity values of 768 analytes within 24 hours. Specialized system software includes a hit selection feature for normalization, merging, and analysis of fragment or other screening data sets to accurately select hits.
- Simplified kinetic characterizations - With OneStep injections, users can obtain accurate kinetics and affinity values from a single analyte concentration injection eliminating the need to run multiple analyte titrations; saving time and reducing errors associated with preparing serial dilutions. NextStep injections provide a simple workflow to characterize competitive binders against a ligand to capture the mechanism of action.
- Low baseline noise, minimal drift, high sensitivity and up to 40 Hz data collection rate - Extremely low baseline noise and drift enable measurement of reliable kinetics and affinity values for high affinity and challenging binders. With up to 40 Hz of data acquisition capabilities, Pioneer FE systems are capable of capturing very fast kinetic association and dissociation phases of a wide range of biomolecules and small molecule fragments.
Pioneer SPR systems provide high quality kinetic data (ka, kd) and affinity measurements (KD) to characterize a wide range of biomolecular interactions from small molecule fragments to biologics. OneStep SPR injections enable faster assay workflows where a single analyte concentration is sufficient for accurate kinetics and affinity determination.
With three sensing channels, two target proteins or ligands can be analyzed simultaneously for binding with one surface utilized as a reference channel.
Offers a variety of sample vessel formats with up to two 384 well plate capacity with an unattended runtime of about 72 hours for performing large binding screens.
Pioneer SPR systems are equipped with an integrated, industry-proven data analysis software based on Scrubber and Clamp platforms. Raw data across multiple runs, days, and sensors can be combined and analyzed using binding algorithms such as single site, multi-site binding, mass transport, and irreversible inhibitor analysis as required for analysis.
As the analyte gradient is generated in OneStep injections, the diffusion coefficient is calculated that provides an assessment of aggregation enabling users to gain information not only on kinetics and affinity but also on aggregation in just one injection.
The Pioneer fluidics system is made up of inert materials such as PEEK, ceramic and Tefzel providing durability and low maintenance.
Applications
Fragment Screening
Fragment-based drug design has become an increasingly popular platform for the identification of lead candidates in drug discovery programs. The detection and characterization of fragment binding events is facilitated by sensitive biophysical technologies capable of detecting low affinity interactions of low molecular weight compounds.
The Pioneer FE system has the necessary sensitivity and throughput to provide complete fragment screens on libraries of several thousand compounds in just a few weeks per target.
Characterization of High Affinity Biologic Interactions
Target binding characterization is an essential analytical step for the selection of high affinity (KD <1 nM) and highly specific biologics regardless of the types of molecules. A kinetic analysis further describes the components of association and dissociation that comprise the overall affinity interaction. For example, two lead compounds may possess a similar affinity (KD) to its target, but their differences in kinetic rate constants of association and dissociation can be used to estimate which will be more useful in vivo. Accurate analysis of these kinetic rate constants is therefore important information for lead selection and predicting the efficacy of protein therapeutics.
The Pioneer FE system with next generation injections improves the efficiency of the characterization process over traditional SPR by determining the kinetics and affinity in a single step. The next-generation OneStep gradient injection featured on the Pioneer platform dramatically increases the speed of affinity characterization while maintaining accuracy and high confidence in results.
Characterize Irreversible Inhibitors and Measure Commitment to Covalency
The majority of small molecule inhibitor assays tested with label-free, real-time biosensor technologies are reversible interactions, characterized by commonly used kinetic rate models. However, a significant fraction of therapeutic enzyme inhibitors on the market functions through covalent modification of the target.
The Pioneer FE is a SPR platform that can be used with regenerable Streptavidin biosensors to reversibly capture protein targets and quantify the efficiency of covalent inhibitors binding to the target. The Pioneer FE system's irreversible inhibitor applications method can be used to determine inhibitor compound's commitment to covalency (Cc) as a metric for irreversible inhibitors.
Featured Publications
| Refractive index range | 1.33–1.40 |
| Short term noise | < 0.035 RU |
Long term drift | < 0.3 RU/min |
| Molecular weight detection | No lower limit for organic molecules |
| Working ranges | ka 102 – 109 M-1 s-1 kd 10-6 – 2.5 s-1 KD ~10-3 – 10-12 M Concentration ~10-3 – 10-12 M |
| Sample capacity | 2 sample racks, 2 reagent racks |
| Sample configuration | 96 vial, deep well and PCR formats, 384-well microplates, custom high volume |
| Sample temperature control | 4 to 40°C (max 15° below ambient) |
| Sample loading | Automatic |
| Number of channels | 3 |
Flow path | 1, 1–2, 1–2–3, 3, 3–2, 3–2–1 |
| Flow channel volume | < 90 nL |
Channel-channel dead volume | < 20 nL |
Real time reference curve subtraction | Yes |
| Injection volume | 2–700 μL |
Injection rise and fall time | < 0.75 second @ 25 μL/min |
| Simultaneous injections | Yes, dual sample loops |
Gradient injections | Yes, OneStep, NeXtStep |
OneStep injections in high throughput mode | Yes |
| Inline buffer degassing | Yes |
| System temperature control | 4–40°C (Max 15° below ambient) |
| Variable data rate | 1–40 Hz |
| Automation capabilities | > 72 hour unattended operation |
| Qdat hit selection feature | Yes |
- Poster: SensiQ Pioneer Drug Discovery Fragments PDF | 112.4 KB
- Octet® SPR Sensor Chip Selection Guide PDF | 1.1 MB
- Flyer: Next Generation SPR Based Interaction Analysis PDF | 359.6 KB
- Flyer: OneStep Injection Technology SPR Based Screening Characterization PDF | 518.4 KB
- Application Note: Fragment Based Drug Discovery On Pioneer Systems Using Next Generation SPR Analysis PDF | 2.7 MB
- Application Note: Kinetics of Irreversible Inhibitors on Pioneer FE System PDF | 828.7 KB
- Application Note: OneStep Lead Characterization of High Affinity Biologic Interactions With Pioneer SPR Systems PDF | 1.0 MB
