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Two ways to run binding

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Binding screening (2 concentrations)

  • What it is: Fast triage using two target concentrations to sort variants by binding strength.
  • When to use: Early discovery and library down‑selection.
  • Output: Strong / Medium / Weak / Non‑binder classification with ranked responses.

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Affinity characterization (4+ concentrations)

  • What it is: Multi‑concentration series (typically 5–7) with global fitting for precise KD, kon, and koff.
  • Why this works: Multiple concentrations across orders of magnitude constrain the fit, separating association (kon) and dissociation (koff). This yields reliable KD across a wider range (≈0.1 nM to 10 µM) than single‑point reads.
  • When to use: Lead optimization, benchmarking, and mechanism studies.
  • Output: KD, kon, koff with confidence intervals and fit diagnostics.

Expression

Confirm expression and yield before binding.

Thermostability

Use Tm to pick robust variants.

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Choose the right depth

  • Screening: Maximize throughput and minimize cost to find winners.
  • Characterization: Quantify kinetics to pick and improve the best.

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FAQs

Our assay loads your constructs onto sensors and measures association/dissociation against a target analyte. If expression is None/Low, signal-to-noise may be insufficient for precise KD. Stronger expression yields cleaner fits.
Typically ~0.1 nM to 10 µM. At the extremes (very tight/weak) accuracy drops; we can adjust concentrations and contact times to bias for tight or weak interactions as needed.
True: clear binder above controls. False: expresses but at/below control signals. Unknown: no/low expression or marginal signals. Strength buckets: Strong (sub 50 nM), Medium (50 nM–1 µM), Weak (>1 µM).
We record time and wavelength shift at 5 Hz and fit 1:1 models; when multiple concentrations are run, we use global fitting across the series.
Screening uses 1–2 concentrations to triage binders; uncertainty is higher than multi‑concentration characterization designed for precise KD, kon, koff.
Yes. We can run cross‑reactivity and polyspecificity panels (e.g., BVP, BSA/HSA); full‑serum profiling is in development.
We screen new targets for NSB and adjust buffers or suppliers. High NSB can lead to negative‑shift curves after reference subtraction.
May indicate multi‑step mechanisms or conformational change. Relative rankings remain useful; precise kinetics may require expanded concentration series.
Yes. We run neutralization/inhibition and competition formats (IC50, epitope access) on request.
Ready to set up a binding experiment? Visit start.adaptyvbio.com/binding.