Summary: BIS as a Research Tool
3.5.23.
The study of protein immobilization and DNA nonselective capture document the versatility and usefulness of BIS for probing bioligand interactions. Indeed BIS is a novel powerful tool, capable of providing insight into kinetic processes that control capture and release of target analytes on solid supports, and quality of the materials provided by different vendors.
Case Study #1: The discovery of a flaw in a widely used immobilization protocol, that has been designed on the basis of trial and (mostly) error, rather than on monitoring of the immobilization process, confirms the need for automated monitoring of immobilization protocols. In this way, the BIS technique provides warning and allows improvement of different types of biomolecular immobilization protocols.
Case Study #2: Investigation of binding capacity should serve only as example
of an experimental approach, since important variables such as flow rates, amount of solid phase, concentration of ligand to be bound, etc. were not investigated. Yet this example will hopefully inspire others to design a more rigorous approach. Indeed, estimate of binding capacity of ligands that cannot be eluted, such as biotin-streptavidin is not a trivial matter, since so far only indirect methods – elution of a colored marker weakly bound to biotin – have been available.
Case Study #3: It is disappointing to realize how much varies the quality of materials offered by different vendors. This finding should serve as a warning to all of us who tend to rely on manufacturers specifications!
