Proteins can be coupled either covalently or via a specific tag to a sensor surface.

 

In interaction analysis with a potent binding partner we can roughly predict the binding kinetics using only two concentrations of the analyte (qualitative analysis) or we determine the exact kinetic data (kass, kdiss, KD) using 8-10 different concentrations of the binding partner (quantitative analysis).



Isoform-specific binding of PKA regulatory subunits to the catalytic subunit, different association/dissociation kinetics (alpha vs. beta), influence of metal ions and nucleotides on binding kinetics, cooperative effects due to dimerisation (monomer vs. d

1. QUALITATIVE Interaction Analysis : 

  • determine qualitatively the interaction of given binding partners for estimation of the KD-value and of the expected kinetic of the interaction
  • optimise buffer conditions to reduce unspecific binding
  • determine the influence of pH, ionic strength and ligands on the given interaction
  • optimise regeneration conditions including tests of functionality of the immobilised ligand after regeneration
  • investigation of mass transfer effects

2. QUANTITATIVE Interaction Analysis 

  • determine concentration series of analytes, the basis for determining association rate constants
  • calculate association and dissociation rate constants (kass, kdiss) by determining a row of concentrations
  • evaluation of binding data including blank subtraction
  • calculation of KD-values using different algorithms depending on the binding kinetics
  • determination of the formation of multiple protein complexes
  • four parallel flow cells permit simultaneous interaction analysis of several interaction partners

PROTEIN / PEPTIDE INTERACTIONS

Peptide and peptide fragments selected from phage display libaries have been extensively used in studying antigen / antibody interactions and to explore the therapeutic feasibility in drug discovery processes. 

 

Since direct immobilisation of the peptide is not always favorable it becomes necessary to immobilise the protein to the sensor surface for some interaction studies. Biaffin has developed methods to analyze even low response signals of peptides to generate reproducible kinetic data even for this class of compounds.

 

kinetic analysis of peptide inhibitor HT31 binding to PKA RII alpha
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