Synthesis of Biotin-Containing Phosphoramidite Linker with Polyether Spacer Arm

Abstract

A phosphoramidite linker unit, based on glycerol backbone and containing a biotin residue attached through a tetraethylene glycol spacer arm, was synthesized. DMTr-Glycidol and tetraethylene glycol were used as starting materials. After conversion of one of hydroxy groups in tetraethylene glycol into an amino group, the epoxy cycle in DMTr-glycidol was opened by this amino alcohol, resulting in the corresponding ether and some quantity of secondary amine. After attaching of biotin residue to the ether followed by phosphitylation, the desirable linker was obtained. The structure of the linker was confirmed by ¹H-¹H COSY, ¹H-¹³C HSQC, ¹H-¹³C HMBC, ¹H-¹⁵N HSQC, and ¹H-¹⁵N HMBC spectra. The resulted phosphoramidite linker unit is suitable for use in common DNA synthesizers. This approach can be used for preparation of various modifiers containing reporter groups attached to the primary amino function using conventional procedures.

Keywords:

• Biotin
• phosphoramidite
• linker
• polyether spacer arm

Acknowledgments

The authors would like to thank Professor Jouko Vepsäläinen (University of Eastern Finland) and Dr. T. Balashova (Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences) for help with NMR spectra.