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 k_a (M‑1 s‑1)
• Disso­ci­a­tion rate constant k_d (s‑1)
• Equi­lib­rium constant K_D (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)