The aim of the course is to provide students with theoretical and practical knowledge necessary for designing, performing and interpreting biomolecular-interaction experiments using biolayer interferometry. In this flash course we will illustrate the method for influenza A virus-sialoglycan receptor, glycoprotein-lectin and/or antigen-antibody interactions. Bringing your own samples is possible. Please contact us first if you want to do this.
Biolayer interferometry is a label-free technology for measuring biomolecular interactions. It is an optical analytical technique that analyses the interference pattern of white light reflected from two surfaces: a layer of immobilized protein on the biosensor tip, and an internal reference layer (Figure 1). Any change in the number of molecules bound to the biosensor tip causes a shift in the interference pattern that can be measured in real-time.
Two machines are available (OctetQK and OctetRED384). The bait molecule can be bound to biosensors displaying different functionalities (protein A, streptavidin, Ni-NTA, etc) followed by interaction analysis with target molecules using a simple dip-and-read set up in a 96 well plate.
Figure 1. Bio-layer interferometry uses the interference produced from two light reflections of a single source to measure the association/dissociation of a target molecule on the sensor surface. Association or dissociation of the target from the sensor (left) results in a change in wavelength (middle), which is translated into a binding curve (right).
Possible applications include a wide variety of biomolecular interactions, including (but not limited to) interaction of proteins with other proteins/peptides (e.g. antigen/antibody characterization), DNA, RNA, lipids, glycans and small molecules (label-free detection down to 150 Da). Also the interaction of proteins with large assemblies such as (protein-containing) membrane vesicles/liposomes as well as virus-receptor interactions can be analysed (affinity range 1 mM-10 pM).
0.5-1 day in total
Erik de Vries (firstname.lastname@example.org)
Xander de Haan (email@example.com