Ligand Binding Assays

The methods applied in our lab are used to inves­ti­gate binding inter­ac­tions of mole­cules. Resulting para­me­ters include kinetics, affinity, speci­ficity, concen­tra­tion and content. We have broad expe­ri­ence in providing tailor-made appli­ca­tions to help under­stand binding inter­ac­tions in detail.

We offer setup of methods based on quality by design (QbD) prin­ci­ples. Estab­lished proto­cols are avail­able for a wide selec­tion of protein inter­ac­tions (such as Fc receptor binding and C1q binding) and may be adapted to your specific needs. Method qual­i­fi­ca­tion and vali­da­tion is performed in accor­dance with the applic­able guide­lines, e.g. ICH Q2R1.

Surface Plasmon Reso­nance (SPR, Biacore T200)

  • Biomol­e­c­ular inter­ac­tion analysis using a real-time and label-free system

Binding kinetics and affinity

Reportable values:

  • Asso­ci­a­tion rate constant ka (M‑1 s‑1)
  • Disso­ci­a­tion rate constant kd (s‑1)
  • Equi­lib­rium constant KD (M)

Sensor­gram comparison

Reportable value:

  • Simi­larity score (%)
  • Compar­ison of samples to pool” of refer­ence data -> simi­larity corridor
  • Applic­able for complex inter­ac­tions without defined fitting model

Surro­gate potency assays

Reportable value:

  • % rela­tive potency compared to reference
  • Dose-response curve
  • Eval­u­a­tion: PLA, 4PL

Cali­bra­tion-free concen­tra­tion analysis (CFCA)

Reportable value: active concentration

  • Estab­lished assays, e.g.: Fc receptor kinetics and affinity: FcRn, FcγRIa, FcγRIIa, FcγRIIb, FcγRIIIa, FcγRIIIb

ELISA

  • different assay formats, such as direct, indi­rect, sand­wich or compet­i­tive ELISA can be applied
  • Deter­mi­na­tion of rela­tive potency
  • Concen­tra­tion / content of analyte

Estab­lished assays, e.g.: C1q binding (potency), Residual host cell protein (HCP) determination

Glyco­sy­la­tion analysis

  • Lectin array tech­nology (Glyco­Sta­tion)
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