What is the best approach to measuring antibody affinity in _Antibody Technology Column_Technical FAQs_Information Center

Label-free interaction analysis is of great importance for scientists to study interactions between biomolecules. Scientists from Creative Biolabs are competent for performing antibody affinity and kinetics measurement with the most advanced Biacore, ProteOn, Octet systems, etc. These systems are mainly based on the optical phenomena technologies - surface plasmon resonance (SPR) and bio-layer interference (BLI), which enable a direct detection and measurement of molecular interactions in a real-time monitoring manner.

SPR is an extremely sensitive method for the detection of molecular interactions by tracking the change of signal via sensor chips. SPR signal is defined as refractive index changes, which refers the response unit (RU) is approximately equivalent to a surface concentration of protein at 1pg/mm2. Plotting the SPR signal over time during the interaction between two molecules results in a sensorgram, which could be visualized in real time (Figure 1). General binding response includes three phases: association, equilibrium and dissociation; fitting these sensorgram data with a mathematic model allows scientists to calculate the association (Ka) and dissociation (Kd) rate constants and ultimately determine the binding affinity (KD). Different from isothermal calorimetry (ITC) that measures antibody binding affinity based on equilibrium point, SPR could obtain full kinetic parameters to evaluate all the effects of association and dissociation as well as antibody affinity.

Inquired items

Inquired items

All Departments

What is the best approach to measuring antibody affinity in

Online Inquiry