Biologically produced bifunctional recombinant protein nanoparticles for immunoassays

Analytical Chemistry
Anu JääskeläinenMarko Virta

Abstract

Nanoparticles are increasingly used as labels for analytical purposes. In general, nanoparticles need to be functionalized with binding molecules (mostly antibodies or fragments thereof) and label substances using a multistep process that requires several manufacturing and purification steps. Here, we present a biological method of producing functionalized nanoparticles for effective use as label agents in a bioaffinity assay. The particles are based on the globular protein shell of human ferritin. A single chain Fv fragment (scFv) of an antibody is used as the binding moiety and Eu3+ ions as the label substance. Conventional chemical conjugation of the particle and antibody fragment is replaced with genetic fusion between the ferritin subunit and scFv genes. The material, for example, the fusion construct is produced in a single bacterial culture as insoluble forms that are easily purified by centrifugations. The subunits are solubilized and self-assembled, and label ions are introduced by shifting the pH. The functionality of these particles is demonstrated with a bioaffinity assay. This method of producing nanoparticles with inherent antigen binding activity presents several possibilities for the simple production of specifi...Continue Reading

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Citations

Nov 26, 2009·Langmuir : the ACS Journal of Surfaces and Colloids·Annie ChenMatt Trau
Apr 18, 2014·Journal of the Royal Society, Interface·Priyanka BhattacharyaYuehe Lin
Jan 1, 2016·Bioconjugate Chemistry·Makan KhoshnejadVladimir R Muzykantov
Dec 10, 2015·Nanomedicine·Amar Bt Ghisaidoobe, Sang J Chung
Oct 30, 2008·Biotechnology and Bioengineering·Anu JääskeläinenMarko Virta
Jun 3, 2010·Biotechnology Journal·Prabhjyot Kaur DehalDavid John Pritchard
Aug 11, 2011·Nanotechnology·Xiaorong ZhangZhihe Qin
Oct 21, 2016·Chemistry, an Asian Journal·Yu ZhangBrendan P Orner
Feb 17, 2015·Chemical Reviews·Günther JutzAlexander Böker

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