Carterra LSA Platform

Carterra LSA is a versatile, fully integrated, high throughput monoclonal antibody (mAb) characterization platform that combines patented continuous flow microfluidics with array surface plasmon resonance (SPR) detection to deliver up to 100x the data in 10% of the time and with 1% of the sample requirements and consumable costs of other systems.

 

Applications

The LSA enables the following automated applications in single unattended runs:

  • Capture-based kinetics and affinity: Screen up to 1152 mAbs
  • Coupled kinetics: Up to 384 immobilized ligands
  • Epitope binning: Interrogate up to 384×384 mAb/mAb pairwise interactions
  • Quantitation: Measure concentration of up to 1152 mAbs
  • Epitope mapping: Screen up to 384 mAbs against an array of up to 384 peptides
  • Blocking / neutralization assays: 384 assay capacity
  • Regeneration scouting: Scout 96 conditions in parallel
  • General multiplexed applications: Array any combination of up to 384 ligands

Unique Platform Technology

The LSA’s throughput, flexibility and applications are enabled by three core system modules:

 

1.  Patented flow cell technology        2.  Array SPR        3.  Software

1. Unique Patented Flow Cell Design & Sample Deck

The LSA has a unique patented flow cell design that provides both throughput and flexibility via two “modes” that switch automatically depending on the assay format:

Multi-channel mode:
Cartridge consisting of a 96 array for:

  • Continuous flow microspotting for discrete immobilization of up to 96 ligands in parallel
  • Nest up to 4x 96 prints on a single chip surface for 384 interaction measurements in parallel
  • ~200 μL per immobilization

Single-channel mode:
Cartridge covers the entire chip surface for:

  • One-on-many interactions
  • Surface “lawn” preparations
  • ~200 μL per interaction

Multi-channel (96) cartridge

The 96-channel printhead enable a 96 protein array to be immobilized in parallel from flow onto a sensor chip surface in a single step. This can be repeated up to 4 times to build a 384 array on the SPR surface.

The sample deck adds another layer of throughput and flexibility.

96 or 384-well plates can be used in the LSA sample deck with multi-channel access in 3 bays and single-channel access in 2 bays. The sample block has the capacity for 3 x 50ml conical tubes, 5 x 15ml conical tubes, and 8 x 1.5ml tubes.

Automated switching between the single-channel and multi-channel cartridges.

2. Array SPR

A high-resolution CCD camera images the entire chip surface simultaneously, enabling up to 384 real-time interactions to be monitored in parallel with local referencing via neighboring interspots.

3. Industry-leading Data Analysis and Visualization Software

Navigator User Interface

A streamlined user interface enables quick and efficient experimental setup. A diverse range of experiments can be quickly defined using intuitive features, minimizing hands on time prior to assay start.

Hands-on time is further reduced via automated data analysis which enables rapid processing and initial curation of large experiments. Dedicated application-focused analysis software follows intuitive, simple-to-use workflows and powerful visualization tools.

Kinetics Analysis Software

Complete kinetic data processing and analysis can be automated, with referencing, zeroing, cropping, blank subtraction, and kinetic model fitting all executed as a single operation for up to 1152 samples per unattended run.

Epitope Analysis Software

In the binning tool, data are linked across panels to facilitate review of sensorgrams, heat maps, networks, and a dendrogram.

The ability to merge orthogonal data and view networks by color options provides a powerful and intuitive way of assessing your antibody panel’s diversity, while keeping an epitope-centric view.

The network plots above provide an alternate view of the data, where chords represent the blocking relationships, envelopes inscribe the bins, and colors depict merged data.

A binning dendrogram, showing an adjustable custom cut-height (red line at 4.1) next to a community plot that is clustered by the custom cut height, which provides a less granular view than the four network plots above where the cut height was zero.

Available Chip Chemistries

Carterra LSA Sensor Chips

Selection of the appropriate sensor chip surface is essential for robust data quality when designing an Array SPR assay. Carterra offers a broad range of chip surfaces to support the diverse applications available on the LSA. Described here are the standard chip offerings for the LSA. Custom surfaces may be available upon request. Always consult with a Carterra Applications Scientist when selecting sensor chips.

Name Description Application
HC30M Polycarboxylate hydrogel, medium charge density, 30nm coating thickness Moderate ligand capacity and excellent diffusion characteristics
HC200M Polycarboxylate hydrogel, medium charge density, 200nm coating thickness Higher ligand capacity for increased levels of immobilization
CMDP 2D planar carboxymethyldextran surface, <5nm coating thickness Lower ligand capacity and excellent diffusion characteristics
CMD50M Carboxymethyldextran hydrogel, 50nm coating thickness Moderate ligand capacity and improved diffusion characteristics
CMD200M Carboxymethyldextran hydrogel, 200nm coating thickness Higher ligand capacity for increased levels of immobilization
SAD200M Streptavidin, immobilized in a carboxymethyldextran hydrogel, 200nm coating thickness High capacity for biotinylated ligand applications
HCX30M NHS activated polycarboxylate hydrogel, medium charge density, 30nm coating thickness Preactivated HC30M, does not require activation step
HCX200M NHS activated polycarboxylate hydrogel, medium charge density, 200nm coating thickness Preactivated HC200M, does not require activation step

Notes

While CMD is a well described chemistry for use in biosensors, the linear polycarboxylate chemistry of HC and HCX chips provides an alternate surface chemistry to that of CMD in instances of non-specific binding or when differential diffusion characteristics are warranted.

25mM MES pH 5.5, 0.01% Tween-20 is the recommended running buffer for HCX chips in order to extend half-life of the sensor chip surface during coupling of proteins.

CMD and HC/HCX sensor chips can be used to build custom capture surfaces such as anti-mouse or Protein A/G via amine coupling of capture molecules.

When stored at -20 °C CMD and HC/HCX surfaces are good for two years after purchase, while protein derivatized coatings such as SAD should be used within one year.

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