BLI is known as a label-free detection technique that depends on white light reflection from the surface of a biosensor tip.

This technique analyzes how the interference pattern of white light reflected from the tip alters when biomolecules are fixed to it. This shift is quantified in nanometers and is documented in real-time. It changes with the size and number of biomolecules that have been bonded to the tip.

Gator® Pro: High-Level Biolayer Interferometry

Image Credit: Gator Bio, Inc.

Gator® Pro: High-Level Biolayer Interferometry

Image Credit: Gator Bio, Inc.

One Tool. Many Answers.

The Gator® Biolayer Interferometry system examines biological molecules in real time and can be an invaluable tool at various stages of therapeutic development.

The Gator® Pro instrument has been crafted for epitope binning, high-throughput kinetics, and precise quantitation at the time of antibody development.

Also, the device promotes viral vector analytics by examining capsid titers of several AAV serotypes and identifying empty versus full (E/F) content.

The Gator® Pro instrument increases walk-away time while offering rapid, dependable, and high-quality data.

Gator® Pro: High-Level Biolayer Interferometry

Image Credit: Gator Bio, Inc.

Gator® Pro Features

  • Designed for rapid, automated, and high-throughput analysis
  • 32 spectrometers mean that high-frequency parallel measurement of up to 32 samples is possible
  • Three sample plates enable automated data acquisition for 1152 samples for every batch
  • The Gator® Pro system provides precise and high-sensitivity data utilizing Gator Bio next-gen biosensors

Highlights

  • Quick biomolecule characterization
  • Adaptable 3 × 96 or 3 × 384-well format
  • High throughput and high-frequency parallel data acquisition of 32 samples
  • Automated data acquisition and processing of up to 1152 samples for every batch

Gator® Pro: High-Level Biolayer Interferometry

The Gator® Pro instrument accommodates three sample plates of either 96 or 384-well format. A fourth plate is reserved for biosensors and regeneration. Image Credit: Gator Bio, Inc.

Gator® Pro: High-Level Biolayer Interferometry

Image Credit: Gator Bio, Inc.

Epitope Binning

The Gator® Pro system is capable of accomplishing 32 × 32 epitope binning in a classical sandwich assay or a single automated tandem in as little as eight hours.

Gator® Navigator software powers the workflow. Simple data visualization and presentation make interpretation simple. A larger binning matrix can be created by combining multiple assays. The report for one 96 × 96 competition profile can be completed in less than 5 days by integrating eight 32 × 32 binning assays.

Highlights

  • Around 32 parallel competitive reactions can be performed in just 10 minutes
  • Single 32 × 32 EP assay in less than eight hours
  • 96 × 96 report in five days
  • Wide suite of biosensors available for both sandwich and tandem format

Gator® Pro: High-Level Biolayer Interferometry

Three epitope binning formats on Gator® Pro. Image Credit: Gator Bio, Inc.

Gator® Pro: High-Level Biolayer Interferometry

32 × 32 mAb competition matrix performed in less than 8 hours. Image Credit: Gator Bio, Inc.

High Throughput Kinetics

The Gator® Pro system has the potential to identify the kinetics of a drug molecule binding to its target.

For antigen-antibody interactions, dissociation rates (koff), association rates (kon), and equilibrium dissociation constants (KD) can be assessed with or without the use of labeled reagents. The ultra-stable baseline enhances the quality of high-affinity kinetic data even further.

The Gator® Pro system, with 32 high-frequency parallel measurements, enables high throughput primary screening of antibody libraries. A wide range of biosensors and assay configurations can also be used to precisely determine off-rate ranking in crude media and complete binding characterization of a purified antigen-antibody binding pair.

Gator® Pro: High-Level Biolayer Interferometry

Kinetic sensorgrams of 4 different biosimilars in a single run. Image Credit: Gator Bio, Inc.

Highlights

  • High-frequency parallel determination of 32 various binding reactions
  • Analyte concentration ranges that can be tailored for accurate outcomes
  • Rapid binding constant determination can be carried out in as little as 10 minutes
  • An extensive range of biosensors to assist multiple kinetic assay configurations

Gator® Pro: High-Level Biolayer Interferometry

Gator® Pro system enables 32 parallel acquisitions for high-quality screening and pairing readouts. Image Credit: Gator Bio, Inc.

Antibody Quantitation

The Gator® Pro system’s potential to read 8, 16, 24, or 32 wells in parallel enables an adaptable assay design that maximizes analytical throughput.

The 32-biosensor mode enables quick whole-plate quantitation of 96 or 384 samples in as little as 12 or 32 minutes rather than hours.

1152 samples can be examined per batch with the help of just 32 biosensors and on-board regeneration.

Performance

Table 1. Accurate and precise data for 1152 human IgG sample analysis using Gator Bio Protein A biosensors. Source: Gator Bio, Inc.

Known conc.
(μg/mL)
Calculated conc. (μg/mL) Binding rate Standard deviation % CV
(n = 144)
700 704 1.2888 0.0541 4%
300 297.79 0.7435 0.0344 5%
100 103.74 0.297 0.0159 5%
30 28.35 0.0807 0.0045 6%
10 10.14 0.026 0.0012 5%
3 3.19 0.0069 0.0004 6%
1 0.98 0.0016 0.0001 9%

 

Highlights

  • Comes with a 5-log dynamic range for titer determination
  • 32 concurrent biosensors provide unrivaled titer throughput
  • Ready for integration with plate handler for extended walk-away operation
  • Up to 1152 samples for every batch from three sample plates

Gator® Pro: High-Level Biolayer Interferometry

Heat map generated by software for human IgG concentration analysis using Gator Bio Protein A biosensors. Image Credit: Gator Bio, Inc.

Viral Vector Analytics

The Gator® Pro system is able to quickly and accurately determine AAV capsid titer and empty/full ratios.

Diverse AAV serotypes, such as chimerics, can be quantified from crude media and purified samples with the help of a simple “dilute and dip” workflow.

Compared to multiple methods, like PCR, ELISA, and AUC, which tend to be utilized for the same analysis, the high throughput Gator® Pro allows analytics on a single platform.

Performance

Table 2. Dynamic range and reproducibility of AAV9 titer with high sensitivity AAV9 kit. Source: Gator Bio, Inc

Known conc.
(vp/mL)
Calculated conc. Standard deviation % CV
(n=6)
1.00E+09 8.70E+08 3.50E+07 4%
5.00E+08 5.31E+08 4.87E+07 9%
2.50E+08 2.78E+08 1.91E+07 7%
1.25E+08 1.31E+08 6.06E+08 5%
6.25E+07 6.60E+07 2.90E+06 4%
3.13E+07 3.06E+07 2.69E+06 3%
1.56E+07 1.58E+07 5.37E+05 9%
7.80E+06 7.60E+06 3.95E+05 5%

 

Highlights

  • Compatible with all serotypes, including chimerics
  • Up to 32 E/F ratio determinations
  • 96 upstream titer samples completed in 100 minutes
  • “Dilute and dip” workflow for upstream titer determination from complex matrices
  • High throughput crude sample analytics
  • Easily customizable assays

Gator® Pro Specifications

Gator® Pro: High-Level Biolayer Interferometry

Image Credit: Gator Bio, Inc.

General

Table 3. Source: Gator Bio, Inc.

. .
Detection Biolayer Interferometry
Microplate positions 4 total (3 x 96- or 384-well microplates and 1 x Max Plate)
Sample type Proteins, antibodies, peptides, nucleic acids, liposomes, viruses, and small molecules
Maximum sample capacity 1152
Software Integrated
Simultaneous reads 8, 16, 24, and 32
Spectrometers 32
Acquisition rate 2, 5, and 10 Hz
System baseline drift ≤ 0.015 nm/hour
Dimension (H x W x D) 84 x 114 x 77 cm
Weight 220 kg
Automation compatible Yes
Orbital flow 100 – 2000 rpm
Analysis temperature range Ambient plus 4 °C to 40 °C

 

Kinetics

Table 4. Source: Gator Bio, Inc.

. .
Analysis time Real-time kinetic binding from 5 minutes to 4 hour
Baseline noise (RMS) ≤ 4 pm (8-32 biosensors)
Baseline drift ≤ 0.1 nm/hour
Association rate (kon) 101 to 107 M-1 s-1
Dissociation rate (koff) 10-6 to 10-1 s-1
Affinity constant (KD) 1 mM - 10 pM
Molecular weight >150 Da (8-32 biosensors)

 

Quantitation

Table 5. Source: Gator Bio, Inc.

. .
Analysis time 32 samples in 2 minutes, 1152 samples in 142 minutes
Quantitation range (Protein A Biosensor) 0.02 – 2000 μg/mL
Quantitation precision CV < 10%
AAV upstream analysis 96 samples in 100 minutes

 

Epitope Binning

Table 6. Source: Gator Bio, Inc.

. .
Analysis time Single run with 32 x 32 less than 8 hour
Pairing format In-tandem, classical sandwich and pre-mix
Binning capacity 32 x 32

 

Gator® Pro: High-Level Biolayer Interferometry

Image Credit: Gator Bio, Inc.

The patented Gator® solution for biotherapeutics discovery includes BLI instruments, chemistry, biosensors, and software.

Gator® Pro: High-Level Biolayer Interferometry

Image Credit: Gator Bio, Inc.

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