Ultrasensitive immuno-detection using viral nanoparticles with modular assembly using genetically-directed biotinylation

Julia Litvinov, Anna E.V. Hagström, Yubitza Lopez, Meenu Adhikari, Katerina Kourentzi, Ulrich Strych, Federico A. Monzon, William Foster, Philip T. Cagle, Richard C. Willson

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

We report a novel, modular approach to immuno-detection based on antibody recognition and PCR read-out that employs antibody-conjugated bacteriophage and easily-manipulated non-pathogenic viruses as affinity agents. Our platform employs phage genetically tagged for in vivo biotinylation during phage maturation that can easily be linked, through avidin, to any biotinylated affinity agent, including full-length antibodies, peptides, lectins or aptamers. The presence of analyte is reported with high sensitivity through real-time PCR. This approach avoids the need to clone antibody-encoding DNA fragments, allows the use of full-length, high affinity antibodies and, by having DNA reporters naturally encapsulated inside the bacteriophage, greatly reduces nonspecific binding of DNA. We validate the efficacy of this new approach through the detection of Vascular Endothelial Growth Factor, a known angiogenic cancer biomarker protein, at attomolar concentrations in bronchoalveolar lavage fluid.

Original languageEnglish (US)
Pages (from-to)1863-1868
Number of pages6
JournalBiotechnology Letters
Volume36
Issue number9
DOIs
StatePublished - Sep 2014

    Fingerprint

Keywords

  • Antibody recognition
  • Bacteriophage
  • Biomarker
  • Immuno-PCR
  • Immuno-detection
  • Immuno-phage assay
  • Vascular Endothelial Growth Factor

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Litvinov, J., Hagström, A. E. V., Lopez, Y., Adhikari, M., Kourentzi, K., Strych, U., Monzon, F. A., Foster, W., Cagle, P. T., & Willson, R. C. (2014). Ultrasensitive immuno-detection using viral nanoparticles with modular assembly using genetically-directed biotinylation. Biotechnology Letters, 36(9), 1863-1868. https://doi.org/10.1007/s10529-014-1555-9