The Gator Prime system was designed using biolayer interferometry (BLI) technology, which allows for real-time monitoring. This device can be used at different stages of therapeutic development to assist in the analysis of biological molecules.

The Gator instrument helps to quantify the interference pattern generated by fluctuations in protein concentrations localized to the probe tips. This is useful for examining protein–protein interactions, such as determining binding constants, off-rate screening, affinity maturation, yes/no binding to target antigens, and epitope binning. 

Gator® Prime: A biolayer interferometry system

Image Credit: Gator Bio, Inc.

Gator® Prime: A biolayer interferometry system

Image Credit: Gator Bio, Inc.

The GatorPrime System

The GatorPrime system is comprised of instrumentation, software, and a selection of probes. The integrated software combines acquisition and analysis to enable setup from beginning to end. On the GatorPrime system, several ELISA tests, such as antibody titer measurement and isotyping, can be easily implemented.

Table 1. Source: Gator Bio, Inc.

Gator probe Function Application Dynamic range Regeneration
Protein A Binds IgG’s of various species, including human and mouse Q/K/QKR 0.02 – 2000 μg/mL Yes
Protein G Binds IgG’s of various species, including human and rat Q/K/QKR 0.02 – 2000 μg/mL Yes
Protein L Binds IgG’s of various species through the kappa light chain Q/K/QKR 0.1 – 2000 μg/mL Yes
Human Fc Immobilize human Fc-fusion protein or human IgG for quantitative or kinetic analysis Q/K/EP 0.02 – 300 μg/mL Yes
Mouse Fc Immobilize mouse Fc-fusion protein or mouse IgG for quantitative or kinetic analysis Q/K/EP 0.02 – 300 μg/mL Yes
Anti - FAB Binds F(ab), F(ab’)2, Fc receptor, and full length Human IgG Q/K/QKR/EP 0.3 – 3000 μg/mL Yes
Anti – His Binds His-tagged proteins Q/K/QKR/EP Protein-dependent Yes
Streptavidin (SA) Binds biotinylated and Avi-tagged biomolecules Indirect Q/K/EP Protein-dependent No
Small Molecule, Antibody, and Protein (SMAP) Binds biotinylated and Avi-tagged biomolecules and subsequent binding of small molecules and protein Indirect Q/K >150 Da No
APS (Aminopropylsilane) Binds hydrophobic proteins Q Protein-dependent No
Amine-Reactive Covalently attach amine group of proteins using EDC/NHS Indirect Q/K/EP Protein-dependent No
Custom-Made Custom-made biosensors for your specific applications (Ex: SARS-CoV-2 R&D biosensors for COVID research) Varies Varies Varies


Epitope Binning With Gatorprime System

When an antibody binds to an antigen, it does not bind to the complete protein sequence. Instead, the antibody attaches to a segment of the antigen that is approximately five or six amino acids long.

As such, a typical protein contains numerous epitopes to which antibodies bind. When it comes to epitope binning, antibodies are evaluated in a paired combinatorial manner, and those that compete for the same binding area of the antigen are put together into bins.

An epitope bin is a relative concept determined by the epitopes represented in the evaluated mAb panel.

Based on signal fluctuations or nanometer shifts, the GatorPrime system tracks real-time binding and determines whether a certain antigen-antibody pair should be in the same epitope bin.

Gator Bio probes (Mouse Fc, Human Fc, Anti-His, etc.) are available for epitope binning. The experiment can be configured to run in premix, in-tandem, or traditional sandwich forms. The materials and probes can be used multiple times.

Gator® Prime: Biolayer Interferometry

Image Credit: Gator Bio, Inc.

Gator® Prime: Biolayer Interferometry

Epitope binning binding curve using in-tandem assay. Image Credit: Gator Bio, Inc.

Gator® Prime: Biolayer Interferometry

Epitope binning against supernatant. Image Credit: Gator Bio, Inc.

Antibody Titer Using Gator Bio Protein A Biosensors

High-sensitivity Gator Bio Protein A biosensors enable the efficient and specific capture of any Fc-tagged protein that binds to protein A. In most commercially available media, the dynamic range of Gator Bio Protein A biosensors spans from 25 ng/mL to 2 mg/mL. This allows for a wide range of hybridoma supernatants to be investigated using a single setup.

Gator Bio Protein A biosensors may be regenerated up to 20 times under simple regeneration conditions, and the same biosensors can be used for several days.

Gator® Prime: Biolayer Interferometry

Standard curve for human IgG binding to GatorPrime Protein A biosensors. Image Credit: Gator Bio, Inc.

Gator® Prime: Biolayer Interferometry

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

Kinetic Characterization Using the Gatorprime System

The Gator system can also be used to determine the kinetics of a drug molecule interacting with its target. The system enables quick measurements of the dissociation rate (koff), association rate (kon), and equilibrium dissociation constant (KD) of antigen-antibody interactions with or without labeled reagents.

Alternatively, the Gator system can be used for primary kinetic screening of antibody libraries in various crude media to determine off-rate ranking and comprehensive binding characterization of a purified antigen-antibody binding pair.

An extensive range of probes are available for the kinetic characterization of biological samples. In general, Gator Bio biosensors are highly adaptable and regenerable (excluding the SA biosensors) and can be used multiple times. This capability enables the user to establish several real-time binding assays with blank sensors and proper controls to get high-quality binding data.

Gator® Prime: Biolayer Interferometry

Kinetic analysis of a purified antigen-antibody pair using Gator Bio Streptavidin biosensors. Image Credit: Gator Bio, Inc.


  • Eight various binding reactions can be performed at the same time
  • A wide range of biosensor choices to quantify binding kinetics
  • Easy to customize assay and fine-tune concentration ranges of analyte to get precise binding constants
  • The binding constant can be identified within 20 minutes

Antibody Isotyping and Subtyping with Gatorprime System

Immunoglobulins, or antibodies, can be divided into five categories: IgA, IgD, IgG, IgE, and IgM. Within these five primary classes, different species may have different subclasses or isotypes of antibodies.

These subclasses differ in the disulfide links that connect the antibody's two heavy chains in the constant region. For example, mice have IgG1, IgG2a, IgG2b, and IgG3. Gator Bio Human Fc (HFC) and Mouse Fc (MFC) biosensors are commonly used to determine isotypes (IgG1, IgG2a, etc.).

Users are able to definitively verify the presence of antibody subtypes in crude samples, like hybridoma supernatants, thanks to the availability of Gator software.

Gator® Prime: Biolayer Interferometry

Hybridoma supernatant isotyping using Gator Mouse Fc biosensors. Image Credit Gator Bio, Inc.

Gator® Prime: Biolayer Interferometry

96-well plate with unknown isotypes of mouse IgG hybridoma supernatant characterized using Gator Mouse Fc biosensors. Image Credit: Gator Bio, Inc.


  • Antibody isotype determination can be performed even in crude samples
  • Regenerable biosensors to save time and assay or cost
  • High sensitivity
  • Visual real-time binding outcomes


Table 2. Source: Gator Bio, Inc.

Detection Biolayer interferometry
Number of channels 8
Data collection rate 2 Hz, 5 Hz, 10 Hz
Orbital flow 0 – 1,500 rpm
Instrument 460 mm x 670 mm x 320 mm (H x W x D)
30 kg
Electrical 110 V – 220 V 5 A
Power 200 W



  • Smart search and a preview of outcomes
  • Enables initiation of the next assay while the present one is in progress
  • QKR — quantitation, kinetics, and regeneration in one run
  • Real-time display of quantitative outcomes and heat map
  • Operating system: Windows 10 Professional 64-bit
  • Combined data acquisition and analysis
  • Data analysis templates available for multiple binding models
  • Epitope binning setup and analysis


Table 3. Source: Gator Bio, Inc.

Sample types Proteins, antibodies, peptides, DNA, RNA, liposomes, cells, viruses, etc.
Work flow
  • 2 x 96-well plates, in tilted or flat position
  • Up to 8 samples in parallel, up to 96 samples per 96-well plate
Types of analysis
  • Yes or no binding
  • Relative and absolute concentration
  • Kinetics and affinity



Table 4. Source: Gator Bio, Inc.

Analysis time
  • IgG concentration in 30 seconds for 8 samples
  • <10 minutes for 96 samples
Linear range for Gator Protein A biosensor 0.025 – 2,000 μg/mL



Table 5. Source: Gator Bio, Inc.

Analysis time Real-time kinetic binding from 1 minute to 4 hours
Association rate kon 101 – 106 M-1 /s
Dissociation rate koff 10-6 – 10-11 /s
Affinity constant KD 10 pM – 1 mM
Binding curve baseline noise <3 pm (RMS)
Binding curve baseline drift <40 pm/hour (RMS)

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