Octet® RED96e System

Octet RED96e System

The new Octet RED96e system is an enhancement to our industry-leading Octet RED96 instrument with additional capabilities to expand the number and types of samples that can be assayed, increasing access to data that can reduce the risk in bringing a drug candidate to market.

From screening biologics candidates to uncovering new insights into biophysics and cellular signaling, Octet RED96e system is your partner in finding the next big thing faster. Using Bio-Layer Interferometry (BLI) to measure binding interactions of small molecules, proteins, antibodies, and even cells, determine specificity, calculate titer, characterize affinity, and more. Assays can now be performed over a wider temperature range of 15-40°C on the Octet RED96e system, allowing for kinetics measurement of unstable proteins at lower temperatures, or biologically relevant molecules at physiological temperatures. An evaporation cover for microplates also allows for greater walk-away run time, with minimal sample evaporation for up to 12 hours. The new Octet Data Analysis HT software version released with the Octet RED96e system brings custom reporting options and multi-experiment analysis that reduces analysis time from hours to minutes.

Key Benefits

  • Eight parallel, independent channels for maximum speed and assay flexibility
  • Versatility to detect biomolecules from small molecules up to mammalian cells
  • High-quality kinetic screening and affinity characterization for a wide variety of biomolecules. Measurement of long dissociation times possible for multiple high affinity binding pairs.
  • Large quantitation dynamic range
    • Simple one-step assay to measure 25 ng/mL to 2 mg/mL of IgG concentration in crude matrices in as little as two minutes per samples or 32 minutes for a whole 96-well plate
    • High sensitivity to sub-ng/mL levels possible with 2-step and 3-step assay formats, allowing automated measurement of contaminants such as host cell proteins and residual protein A
  • Microfluidics-free Dip and Read format reduces assay time and maintenance cost. Combined with biosensor regeneration, the Octet RED96e system’s re-racking option provides ultra-low cost per test.
  • Non-destructive sampling allows full sample recovery. Microplate evaporation cover further minimizes losses in sample volume, allowing post-analysis recovery of precious samples after a 12-hour run.
  • Up to 12 hours of unattended run time. Increased throughput for screening and characterization purposes.
  • Sample plate cooling for temperature-sensitive proteins and for thermodynamic analysis. Determine binding rate constants at multiple temperatures to calculate thermodynamic parameters.
  • New version of Octet Data Analysis HT software enables multi-experiment analysis and customized reports

Quantitation Assays

The Octet RED96e system directly measures the presence of specific proteins and other molecules in solution with minimal interference from complex matrices. Accurate and reproducible concentrations can be determined in as little as two minutes per sample or 32 minutes for a whole plate using a simple, one-step assay (Figure 1). High sensitivity in quantitation can be achieved to sub-ng/mL levels with 2-step and 3-step assay formats, allowing automated measurement of contaminants such as host cell proteins and residual Protein A faster and more precisely than ELISA. Process economics can be improved further by regenerating and re-using the biosensors.

Octet RED96 concentration anaylsis data

Figure 1: Human IgG Quantitation. Example data from human IgG analyte binding to Protein A biosensors. Binding was performed at 30°C, with a shake speed of 1000 rpm and a two-minute read per well. Human IgG solution was prepared at 0.025 µg/mL up to 300 µg/mL and the standard curve shown on a log-log scale was generated using the initial slope algorithm and fitted with the unweighted 5-parameter logistic (5PL) regression model.

Kinetic Assays

The Octet RED96e system monitors binding events in real time to calculate on rates (ka), off rates (kd), and affinity constants (KD). The superior sensitivity of the system enables measurement of small organic molecules (Figure 2) and kinetic constants over a broad range. The temperature of one 96-well sample plate can be controlled from 15-40°C, which enables reliable kinetic determinations from low (Figure 3) up to physiological temperatures for temperature-sensitive proteins. Additional advantages afforded by sample cooling include the ability to rapidly determine binding rate constants at multiple temperatures to extrapolate thermodynamic measurements. The Octet RED96e system’s eight channels can be used independently to measure samples for screening purposes or in tandem, pairing the sample read with a dedicated reference for high-quality kinetic characterization.

Octet RED96 kinetic anaylsis data

Figure 2: Small molecule kinetics. Example data from benzenesulfonamide (MW 157 Da) binding to biotin-carbonic anhydrase loaded on Super Streptavidin biosensors. Binding was performed at 25°C, with a shake speed of 1000 rpm. A 100 µM benzensulfonamide solution was prepared and serially diluted 1:4.

Octet RED96 kinetic anaylsis data

Figure 3: Large molecule characterization. Example data from human Prostate Specific Antigen (PSA, MW 30 kDa) binding to a biotinylated anti-human PSA mouse monoclonal antibody loaded onto Streptavidin biosensors. Binding was performed at 15°C with a shake speed of 1000 rpm. A 200 nM PSA solution was prepared and serial diluted 1:2 to obtain the five concentrations run.