BioLayer Interferometry (BIL) is a label-free optical analysis technique for measuring biomolecular interactions. Our Octet® RED96 System instrument can be used to monitor the relationship between an immobilised protein on the biosensor tip and a solution analyte. Typical analytes (substrates) include proteins, carbohydrates and other small molecules. Results are obtained in real-time through the simultaneous monitoring of a large number of interactions in an automated manner, to generate information regarding protein:substrate binding and kinetics.
The BioLayer Interferometer is ideally suited to measuring the interactions between an immobilised protein and your chosen analyte (substrate). These analytes include proteins, antibodies, peptides, serum-containing media, buffers, crude cell lysates, periplasmic fractions, cell culture supernatant. It can also be used to probe new drug entities, carbohydrates and synthetic polymer interactions.
Samples do not need to be labelled in any way.
Kinetic and affinity analysis (kobs’, ka’, kd’ and KD) between the protein-functionalised biosensor and your analyte of interest. Concentration monitoring and concentration determination.
Data is presented both as plots displaying kinetic binding data, equation fits and residuals of fits and also as tabulated data.
How does it work?
BIL is an optical analysis technique. It analyses the interference pattern of white light being reflected from the immobilised protein layer present on the biosensor tip and the internal reference layer. When changes occur to these layers (such as binding of analyte molecules) the optical thickness of the layers is altered and there is a change in the interference pattern of the light. The shift in the wavelength is directly related to the change in the thickness of the biological layer, allowing binding specificity, association/dissociation rates and concentration to be accurately monitored in real time.
Sample handling requirements:
Analyte kinetic analysis can be performed on a large range of samples, from 150 Da fragments through to large macromolecules such as IgG. Biomolecule concentration analysis can be performed on solutions ranging from ng/ml to mg/mL.
In all cases, sample volumes of 180-220 µL are required. The testing is non-destructive. Up to 8 assays can be run in parallel, with up to 96 samples in each 96-well plate.
Octet® RED96 System.
Dr Ian Hancox, 024 76 150 380
email i dot hancox at warwick dot ac dot uk.
Typical results format, and sample:
|Warwick collect/analyse data|
|Warwick collect data|