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.
Image Credit: Gator Bio, Inc.
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.
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
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.
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
Three epitope binning formats on Gator® Pro. Image Credit: Gator Bio, Inc.
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.
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 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
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
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 |
Image Credit: Gator Bio, Inc.
The patented Gator® solution for biotherapeutics discovery includes BLI instruments, chemistry, biosensors, and software.
Image Credit: Gator Bio, Inc.