Exploring minimal biotinylation conditions for biosensor analysis using capture chips

Papalia, Giuseppe and Myszka, David Exploring minimal biotinylation conditions for biosensor analysis using capture chips. Analytical Biochemistry403: 30-35; (2010). Goto reference

Using Biacore's new regenereteable streptavidin capture (CAP) sensor chips, a number of biotinylation conditions for target ligands were investigated. For instance the use of the standard amine as well as the less commonly used carboxyl biotinylation methods, the time scales required for efficient biotinylation as well as the hazards of overbiotinylation. In Addition a range of desalting methods, including spin columns, dialysis membranes, and filters is tested to remove excess biotin. Finally, the effects of common buffer components, such as Tris and glycerol, on the biotinylation process are tested. Together, this publication can serve as a general guide of the steps to consider when minimally biotinylating a target ligand.

Hahn, C. D. et al Self-assembled monolayers with latent aldehydes for protein immobilization. Bioconjugate Chemistry18: 247-253; (2007).

Aldehyde functions are widely used for immobilization of biomolecules on glass surfaces but have found little attention for biofunctionalization of self-assembled monolayers (SAMs) on gold, due to interference between thiol and aldehyde functions. This problem was recently solved by synthesis of an alkanethiol that carried a vicinal diol group [Jang et al. (2003) Nano Lett. 3, 691-694]. In this article, a general modular strategy was introduced by which SAM components with vicinal diol functions were rapidly synthesized with high yield.

Kyprianou, Dimitris et al New reactive polymer for protein immobilisation on sensor surfaces. Biosensors and Bioelectronics24: 1365-1371; (2009). Goto reference

A novel protocol for the covalent immobilisation of biomolecules containing primary amines using an inexpensive and simple polymer is presented. The reactivity of the polymer is based on the reaction between primary amines and thioacetal groups included in the polymer network. These functional groups (thioacetal) do not need any further activation in order to react with proteins, making it attractive for sensor fabrication. The novel polymer also contains thiol derivative groups (disulphide groups or thioethers) that promote self-assembling on a metal transducer surface.