Unit

UNIT 12.7 Synthesis of Covalent Oligonucleotide-Streptavidin Conjugates and Their Application in DNA-Directed Immobilization (DDI) of Proteins

  1. Ron Wacker1,
  2. Christof M. Niemeyer2

Published Online: 1 OCT 2005

DOI: 10.1002/0471142700.nc1207s22

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Wacker, R. and Niemeyer, C. M. 2005. Synthesis of Covalent Oligonucleotide-Streptavidin Conjugates and Their Application in DNA-Directed Immobilization (DDI) of Proteins. Current Protocols in Nucleic Acid Chemistry. 22:12.7:12.7.1–12.7.15.

Author Information

  1. 1

    Chimera Biotec GmbH, Dortmund, Germany

  2. 2

    Universität Dortmund, Fachbereich Chemie, Dortmund, Germany

Publication History

  1. Published Online: 1 OCT 2005
  2. Published Print: SEP 2005

Abstract

Semisynthetic DNA-streptavidin conjugates are synthesized by covalent coupling of thiol-modified DNA oligonucleotides and streptavidin (STV). The resulting conjugates have binding capacities for four equivalents of biotin and a complementary nucleic acid sequence. The conjugates are purified to homogeneity by ultrafiltration and chromatography, and are characterized by spectrophotometry and gel electrophoresis. Subsequently, the conjugates are applied as molecular connectors in the DNA-directed immobilization (DDI) of biotinylated antibodies using DNA microarrays as immobilization matrices. The results are protein microarrays that can be used for the multiplexed detection of various antigens.

Keywords:

  • biotin-streptavidin interaction;
  • DNA conjugation;
  • bioconjugate purification;
  • electrophoretic characterization of DNA-protein conjugates;
  • protein biotinylation;
  • DNA-directed immobilization;
  • DNA and protein microarray technology