Technology Platform: Fluoresence Polarisation


Our product quantification products are based on fluoresence polarisation technology which can be found on a plate reader. Fluoresence polarisation simplifies product titer measurement, glycosylation profiling, HCP and aggregation detection, eliminating the need for complex, expensive alternatives. 

 

1) The Fluoresence Polarisation plate is coated with a fluorescently labelled IgG-binding peptide. 2) When the IgG sample is added, it binds to the fluorescently labelled peptide 3) Binding is detected using fluoresence polarisation

Our product quantification products are fully automatable, thereby allowing fully scalable workflows. The assay is based on the principle that small molecules rotate faster than larger molecules and the rotation rate can be determined by fluoresence polarisatoin to measure product titre with exceptional ease and reproducibility.

Fluorescence Polarization Technology

Fluorescence polarization (FP) effectively analyses the size of complexes by measuring how fast they rotate in solution i.e. rotational diffusion. When fluorophores are excited by polarised light, they preferentially emit light in the same plane of polarisation when they are “fixed”. However, the rotation of the molecules in the time between absorption and emission of the photon has the effect of “twisting” the polarisation of the light. The more the molecules move in this time, the more the light is depolarised. As a result of small molecules “tumbling” faster in solution than larger molecules, the size of a molecule, with an associated fluorophore, can be measured using the degree of light depolaristion. When the ValitaTITER fluorescently labelled probe is unbound, it tumbles rapidly and depolarises the light more than when it is bound to an IgG, or Fc-region containing protein, which is 5x larger. This change in polarisation can be used to measure the degree of probe binding and thus the amount of antibody in the solution.