Reversible association/dissociation reaction of avidin on the dendrimer monolayer functionalized with a biotin analogue for a regenerable affinity-sensing surface

Abstract

Anew approach to a repeatedly regenerable affinity-sensing surface was developed based on the reversible association/dissociation reactions between avidin and biotin analogues. For the affinity surface, a fourth generation poly(amidoamine) dendrimer monolayer was first constructed on the 11-mercaptoundecanoic acid self-assembled monolayer on gold. The dendritic surface amine groups then were functionalized with biotin analogues, desthiobiotin (1), or newly synthesized desthiobiotin amidocaproate (2), an extended form of 1, which shows lower affinity toward avidin. To test the association/dissociation reaction cycles at the affinity surface, avidin adlayer was formed onto the biotin analogue functionalized surface and displaced with free biotin. To trace the stepwise reactions, biotinylated glucose oxidase (b-GOx) as a model enzyme was loaded onto the affinity surface, and cyclic voltammetric measurements were performed by registering the activity of the associated b-GOx. The efficient association/dissociation reaction cycles were registered, especially for the a-modified electrodes, implying steric hindrance from the ligand length for biospecific interaction. With the optimized affinity-surface construction steps and reaction conditions, continuous association/dissociation reaction cycles were achieved, resulting in a regenerable affinity surface.