Preparation of avidin-labeled protein nanoparticles as carriers for biotinylated peptide nucleic acid

The possibility of preparing protein nanoparticles followed by covalent linkage of avidin was investigated. Free sulfhydryl groups were introduced onto the surface of protein nanoparticles either by aldehyde quenching with cysteine or reaction of free amino groups with 2-iminothiolane. The number of primary amino groups and sulfhydryl groups on the surface of the resulting particles was quantified with site-specific reagents. Avidin was attached to the surface of the thiolated nanoparticles via a bifunctional spacer which reacted in a first step with amino groups of avidin and in a second step with the sulfhydryl groups introduced onto the surface of the nanoparticles. Biotinylated peptide nucleic acid (PNA) as a model compound for biotinylated drugs was effectively coupled to the nanoparticles by complex formation with the covalently attached avidin. Since the formation of the interaction between biotin and avidin is very rapid and stable a highly effective drug carrier system for biotinylated compounds such as PNAs was achieved.