A novel approach to ultrasensitive diagnosis using supramolecular protein nanoparticles

Sung Hyun Lee, Hyewon Lee, Jin Seung Park, Hyoung Choi, Kyung Yeon Han, Hyuk Seong Seo, Keum Young Ahn, Sung Sik Han, Yunjung Cho, Kee Hyoung Lee, Jeewon Lee

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

We report on the ultrasensitive protein nanoprobe system that specifically captures disease marker (autoantibodies of Type I diabetes in this case) with attomolar sensitivity. The system relies on supramolecular protein nanoparticles that bind a specific antibody [65 kDa glutamate decarboxylase (GAD 65)-specific autoantibody, i.e., the early marker of Type I diabetes]. The ultrasensitive detection of early marker of Type I diabetes during the early phase of pancreatic β-cell destruction is important because individuals at high risk of developing Type I diabetes can be identified several years before the clinical onset of the ailment. The bacterial expression of chimera genes encoding N-[human ferritin heavy chain (hFTN-H)]::[specific antigenic epitope]-C produces supramolecular nanoparticles with uniform diameters (10-15 nm), owing to self-assembly activity of hFTN-H. Each nanoparticle, formed by intermolecular self-assembly between the chimera protein molecules, is subjected to carrying a large number (presumably, 24) of epitopes with a homogeneous and stable conformation per autoantibody binding, thereby allowing substantial enhancement of sensitivity. The sensitivity was finally boosted to 3 attomolar concentration of the autoantibodies, 4-9 orders of magnitude more sensitive than conventional immunoassays. Also, this ultrasensitive protein nanoprobe successfully detected natural autoantibodies in the sera from Type I diabetic patients. The attomolar sensitivity was successfully reproduced on the detection of other antibodies, i.e., monoclonal antibodies against hepatitis B surface antigen. With the two antibody markers above, the feasibility of simultaneous and multiplexing-mode detection was also demonstrated.

Original languageEnglish
Pages (from-to)1324-1334
Number of pages11
JournalFASEB Journal
Volume21
Issue number7
DOIs
Publication statusPublished - 2007 May 1

    Fingerprint

Keywords

  • Attomoloar sensitivity
  • Human ferritin heavy chain
  • Nanoprobe system

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this