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Issue 24, 2016
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Design of luciferase-displaying protein nanoparticles for use as highly sensitive immunoassay detection probes

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Abstract

In this study, we developed a protein nanoparticle-based immunoassay to detect cancer biomarkers using a bioluminescent fusion protein. This method relies on the use of protein nanoparticles comprised of genetically-engineered elastin-like polypeptides (ELPs) fused with poly-aspartic acid tails (ELP-D), previously developed in our lab. The sizes of the self-assembled ELP-D nanoparticles can be regulated at the nanoscale by charged repulsion of the poly-aspartic acid chains. To improve the sensitivity of enzyme-linked immunosorbent assays (ELISAs), we herein demonstrate the multivalent display of NanoLuc® (Nluc) luciferase and a biotin acceptor peptide (BAP) on the surfaces of ELP-D nanoparticles, and demonstrate the sensitivity of these multivalent nanoparticles as detection probes. The fusion protein comprised of ELP-D and Nluc-BAP (ELP-D-Nluc-BAP) was found to form nanoparticles with Nluc and BAP displayed multivalently on their surfaces. Moreover, the use of the nanoparticles in ELISA resulted in a detection sensitivity for α-fetoprotein (AFP) about 10 times higher than that of an assay relying on the use of the monomeric version of the fusion protein. Taken together, ELP-D-based nanoparticles displaying multivalent luciferases on their surfaces enable the construction of an ELISA with enhanced sensitivity.

Graphical abstract: Design of luciferase-displaying protein nanoparticles for use as highly sensitive immunoassay detection probes

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Publication details

The article was received on 01 Jun 2016, accepted on 01 Nov 2016 and first published on 02 Nov 2016


Article type: Paper
DOI: 10.1039/C6AN01253A
Analyst, 2016,141, 6557-6563
  • Open access: Creative Commons BY-NC license
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    Design of luciferase-displaying protein nanoparticles for use as highly sensitive immunoassay detection probes

    Y. Ikeda, Y. Mashimo, M. Mie and E. Kobatake, Analyst, 2016, 141, 6557
    DOI: 10.1039/C6AN01253A

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