Antibody Characterisation

Kinetic information about your antibody of interest instead of equilibrium binding data (ELISA)

Classical methods for characterisation of antigen - antibody interactions like ELISA only provide equilibrium binding data at one defined time point not considering rates of complex formation and complex dissociation.
SPR technology has been proven to be an excellent method to validate antibodies by monitoring binding kinetics between antigen and antibody in real time. The obtained binding curves contain detailed kinetic information about association and dissociation rates, about stoichiometry of binding and about binding activity and quality of samples. Furthermore, an assessment of binding characteristics regarding specificity, heterogeneity and avidity effects is feasible as well. Mode of action, temperature dependency and thermodynamic parameters like enthalpie and entropie representing driving forces of an interaction can be examined by analysing antibody antigen interactions at varying temperatures.

Antibody characterisation using SPR technology

Applications of SPR technology for antibody validation are:

  • Qualitative ranking of crude antibody samples from sera, cell lysates or cell culture supernatants for a rapid comparison of binding signals and off-rates
  • Semi-quantitative measurements of antibody antigen interactions in single cycle kinetics for rapid assessment of kinetic rate constants for association and dissociation
  • Quantitative kinetic characterisation yielding kass, kdiss and KD values at high accuracy and reproducibility to obtain a comprehensive kinetic profile of your antibody
  • Pair-wise epitope-mapping to define distinct and overlapping binding epitopes of antibodies
  • Testing cross reactivity of antibodies against similar antigens or antigens from different species
  • Optimisation of experimental conditions for antibody based assays
  • Quality control of recombinant antigen preparations by binding assays
  • Testing immunoreactivity of pharmaceutical compounds in animal samples
  • Quantitative determination of antibody concentrations in serum samples
  • Identification of antibody classes and subtypes (IgG, IgE, IgM, IgG1,...)
  • In-process and quality control during antibody production
  • Stability measurements during different assay conditions (based on concentration determination)
  • Detection and quantification of antibody markers in diagnostic applications
BIAFFIN GmbH & Co KG | Heinrich-Plett-Str. 40 | D-34132 Kassel | Germany
Tel.: +49 (0) 561-804 4661 | Fax: +49 (0) 561-804 4665 |