Vendor-Specific Tips
Practical guidance for the major biosensor platforms. Learn the quirks before they learn you.
π΅ Cytiva Biacore (SPR)
Instruments
- 1 series (1S / 1K / 1S+ / 1K+): Current generation, replacing the T200/8K line in Cytiva's catalogue. 1S/1S+ are 4-flow-cell sensitivity-focused; 1K/1K+ are higher throughput.
- T200: Long-running workhorse for kinetics. 4 flow cells, excellent temperature control. High sensitivity tier.
- 8K / 8K+: High-throughput, 8 needles, faster cycle times.
- S200: High-sensitivity 4-flow-cell instrument positioned alongside the T200 (not entry-level).
- X100: Compact 2-flow-cell instrument, still common in academia.
- 3000 / 4000: Legacy multi-flow-cell systems still in use in many labs.
Software: Biacore Insight Evaluation (1-series, 8K, T200) / BIAevaluation (legacy). Many academic labs also use Scrubber / Scrubber2 (BioLogic Software) as a third-party kinetic fitting tool.
Export formats: .blr (binary), Excel export, sensorgram CSV
Tips & Gotchas
1. Prime Before You Run
Always run a Prime cycle after overnight standby. Biacore flow systems collect micro-bubbles; priming purges them.
2. Normalize Early
Run a Normalize procedure weekly (or after chip changes). This calibrates the detector response across all channels.
3. Watch Your Flow Rate
- Kinetics: 30 Β΅L/min is standard
- Screening: Can go higher (50-100 Β΅L/min)
- If you see mass transport, increase flow rate
4. Reference Subtraction
- Use Fc1 as reference (no ligand) when possible
- Always subtract both reference AND blank injections (double referencing)
5. Regeneration Scouting
Biacore has built-in regeneration scouting wizards. Use them. Common conditions:
- 10 mM Glycine pH 2.0-3.0 (proteins)
- 1 M NaCl (ionic interactions)
- 50 mM NaOH (stubborn binders)
6. CM5 vs CM7 vs NTA
- CM5: General purpose, amine coupling
- CM7: Higher capacity, same chemistry
- NTA: His-tag capture, reversible
π’ Sartorius Octet (formerly ForteBio) (BLI)
Instruments
- R2 / R4 / R8: Current entry / mid-range Octet R series, 2 / 4 / 8 channels respectively. R8 replaces the legacy RED96e.
- RED96 / RED96e / RED384: Legacy 8-channel platforms. Both RED96 and RED384 use 8 channels β what differs is the accessible plate format (96-well vs 384-well), not the channel count.
- HTX: 16-channel high-throughput platform, accesses 96- or 384-well plates.
- SF3: Current high-end successor sitting above the HTX in the Sartorius lineup.
Software: Octet Analysis Studio (current) / Data Analysis HT (legacy)
Export formats: .frd (raw), Excel export, CSV
Tips & Gotchas
1. Hydrate Your Sensors β οΈ
Minimum 10 minutes in buffer before use. This is non-negotiable. Dry sensors = bad data.
2. Orbital Shake Speed
- 1000 rpm for kinetics (standard)
- Lower speeds cause mass transport issues
- Document this in your methods β reviewers ask
3. Black Plates Reduce Noise
Use black 96-well plates to minimize optical interference. Clear plates can introduce artifacts.
4. Signal Saturation / Linear Range
At high shifts (often a few nm, but instrument- and chemistry-dependent), the optical response becomes non-linear with bound mass. Keep loading and binding responses within the linear range. Note: this is signal saturation, not the hook effect β the hook effect specifically describes a signal decrease at very high analyte concentrations in sandwich/immunoassay formats and is a separate phenomenon.
5. Sensor-to-Sensor Variability
Every sensor tip is slightly different. Always run duplicates. Report average Β± SD.
6. Common Sensor Types
| Sensor | Use Case |
|---|---|
| SA | Biotinylated ligands |
| SAX | High-capacity streptavidin |
| AHC | Human IgG Fc capture |
| AHQ | Same as AHC, higher capacity |
| AMC | Anti-mouse IgG Fc capture |
| HIS1K | High-affinity anti-penta-His capture; effectively irreversible |
| NTA / Ni-NTA HC | Ni-NTA capture of His-tagged proteins; reversible, lower affinity than HIS1K |
| Protein A / G / L | Antibody / Fab capture across species |
| AR2G | Amine coupling |
| FAB2G | Fab capture |
π Gator Bio (BLI)
Instruments
- Gator Prime: 8-channel BLI platform
- Gator Plus: 16-channel, higher throughput
Software: Gator Bio Analysis Software
Key Points
Lower Cost BLI Alternative
Gator Bio offers a more affordable entry point into BLI. Capital and consumable costs are typically lower than Octet, but the installed base is smaller β fewer protocols and shared experience to draw on in the literature.
Sensor Tip Compatibility
Uses dip-and-read tips with a similar form factor to Octet. Functionally similar chemistries are available (SA, anti-His, Protein A, etc.), but the underlying surface chemistry, lot QC, and binding capacity are not necessarily identical β validate transfers rather than assuming drop-in equivalence.
Software Differences
Gator's analysis software is its own package and not file-format compatible with Octet Analysis Studio. If you switch platforms mid-project, plan for re-fitting rather than re-loading existing analysis sessions.
π§ Other SPR / BLI Vendors Worth Knowing
These platforms come up in the literature and in lab decisions even though they are less common than Biacore or Octet. Worth knowing exists.
Bruker Sierra SPR
Multi-channel SPR (e.g. Sierra SPR-32 / SPR-24 Pro). Used for higher-throughput kinetics and screening; common in pharma fragment workflows.
Reichert (Reichert4SPR / 2SPR)
Common in academic labs, especially for small-molecule and fragment kinetics where the open fluidics and sensor flexibility are useful.
Horiba OpenPlex
SPR imaging (SPRi) platform β array-format, useful for parallel ligand screening and interaction mapping.
Bionavis (MP-SPR)
Multi-parametric SPR with multi-wavelength capability β gives layer thickness and refractive index in addition to standard kinetics.
AffinitΓ© Instruments (P4SPR)
Compact / portable SPR designed for benchtop or field use. Lower sensitivity than research-grade Biacore but usable for screening.
ITC / DSC
For thermodynamics rather than kinetics: Malvern Panalytical MicroCal (PEAQ-ITC, PEAQ-DSC) and TA Instruments Affinity ITC / Nano DSC are the two main vendors.
π― Which Platform Should I Choose?
Rough decision guide. These are starting points, not absolutes β the right answer depends on sample, throughput, and what's already on your bench.
| Use Case | Reasonable First Choice |
|---|---|
| Small molecules / fragments | Biacore 1-series / T200 / S200 with CM5; Reichert; Bruker Sierra SPR; Creoptix WAVE (waveRAPID for fragments) |
| High-throughput antibody screen | Octet HTX / SF3; Carterra LSA / LSAXT |
| Epitope binning | Carterra LSA (Epitope module) β purpose-built for all-vs-all |
| Precious / scarce sample | Creoptix WAVE (low microfluidic volume); Nicoya Alto (small sample loops) |
| Crude lysate / sticky sample | BLI (Octet, Gator) β dip-and-read tolerates particulates better than microfluidic SPR |
| Teaching / first SPR | Nicoya OpenSPR; Reichert2SPR |
| Thermodynamics (ΞH, ΞS, n) | Malvern MicroCal PEAQ-ITC; TA Instruments Affinity ITC |
π£ Carterra LSA (SPR Array)
Overview
- LSA: 384-ligand array format. Integrated printhead delivers ligands to the SPRi sensor surface. (CFM / Continuous Flow Microspotter is the legacy Wasatch Microfluidics term β current Carterra documentation refers to the printhead.)
- LSAXT: Newer extended-throughput Carterra platform building on the LSA workflow.
- Ideal for epitope binning and large all-vs-all screens.
Software: Carterra Navigator suite, including the Kinetics and Epitope (binning) modules.
Tips & Gotchas
1. Think in Arrays
Carterra excels at "all vs all" experiments. Design your ligand array to maximize pairwise comparisons.
2. Printing Uniformity
Printhead deposition quality matters. Run a QC binding step after printing to verify spot uniformity across the array.
3. Lower Sensitivity per Spot
Each spot is smaller than a Biacore flow cell β expect lower absolute RU values. This is normal.
4. Data Volume
384 spots Γ multiple analytes = huge datasets. Plan your data management strategy before you start.
π Creoptix WAVE (GCI)
Overview
- Grating-Coupled Interferometry
- WAVEsystem with microfluidic cartridges
- 4-channel or 8-channel options
Different Physics, Similar Output
GCI measures the same binding events as SPR but via waveguide interference. Data analysis is analogous.
Cartridge-Based
No chip docking β just snap in a cartridge. Convenient but higher consumable cost per sample.
Low Sample Consumption
Microfluidic channels use very little volume. Good for precious samples.
π· Nicoya (Localized SPR)
Instruments
- OpenSPR: Benchtop, affordable
- Alto: Higher throughput
- Uses gold nanoparticles (LSPR) rather than gold film
Lower Cost of Entry
Good for teaching labs or preliminary screens before moving to Biacore.
Sensitivity Differences
LSPR has a shorter evanescent decay length (tens of nm) than propagating SPR (~150β200 nm). In practice this makes LSPR relatively more sensitive to surface-proximal binding and less sensitive to bulk refractive-index shifts, so absolute response values will differ from a Biacore-style instrument.
Software
Nicoya provides its own acquisition software, and integrates with TraceDrawer (a third-party kinetics analysis package from Ridgeview Instruments, not Nicoya) for kinetic fitting. Good for learning SPR principles.
π Cross-Platform Tips
When Publishing, Always Report:
- β Instrument model and software version
- β Sensor/chip type
- β Temperature
- β Flow rate (SPR) or shake speed (BLI)
- β Buffer composition
- β Regeneration conditions
Comparing SPR vs BLI Data
- ka usually agrees well across platforms
- kd is the most platform-sensitive parameter. Both SPR (rebinding at high Rmax, mass transport) and BLI (drift, local rebinding within the unstirred layer at the sensor tip) can perturb apparent off-rates. Well-designed SPR (low ligand density, high flow rate) typically gives the cleaner dissociation β BLI is generally more prone to apparent kd artefacts, not less.
- KD should be similar if both are measured correctly
- Don't directly compare RU (SPR) to nm (BLI)
Troubleshooting Across Platforms
| Symptom | SPR Likely Cause | BLI Likely Cause |
|---|---|---|
| Linear association | Mass transport, high Rmax | Low shake speed, high loading |
| Unstable baseline | Temperature, ligand loss | Poor hydration, bubbles |
| No binding | Wrong buffer, inactive protein | Same + sensor type mismatch |
| Noisy signal | Air bubbles, pump issues | Plate vibration, sensor damage |