Biologically produced bifunctional recombinant protein nanoparticles for immunoassays

Anu Jääskelainen, Reija-Riitta Harinen, Tero Soukka, Urpo Lamminmäki, Teemu Korpimäki, Marko Virta

Research output: Contribution to journalArticleScientificpeer-review

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. Ibis method of producing nanoparticles with inherent antigen binding activity presents several possibilities for the simple production of specific, functional nanoparticles. Production is fast, economical,and environmentally sustainable, making the system advantageous, particularly in applications requiring large quantities of specific nanoparticles.
Original languageEnglish
JournalAnalytical Chemistry
Volume80
Issue number3
Pages (from-to)583-587
Number of pages5
ISSN0003-2700
DOIs
Publication statusPublished - 2008
MoE publication typeA1 Journal article-refereed

Fields of Science

  • 1182 Biochemistry, cell and molecular biology

Cite this

Jääskelainen, A., Harinen, R-R., Soukka, T., Lamminmäki, U., Korpimäki, T., & Virta, M. (2008). Biologically produced bifunctional recombinant protein nanoparticles for immunoassays. Analytical Chemistry, 80(3), 583-587. https://doi.org/10.1021/ac071382v
Jääskelainen, Anu ; Harinen, Reija-Riitta ; Soukka, Tero ; Lamminmäki, Urpo ; Korpimäki, Teemu ; Virta, Marko. / Biologically produced bifunctional recombinant protein nanoparticles for immunoassays. In: Analytical Chemistry. 2008 ; Vol. 80, No. 3. pp. 583-587.
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Jääskelainen, A, Harinen, R-R, Soukka, T, Lamminmäki, U, Korpimäki, T & Virta, M 2008, 'Biologically produced bifunctional recombinant protein nanoparticles for immunoassays', Analytical Chemistry, vol. 80, no. 3, pp. 583-587. https://doi.org/10.1021/ac071382v

Biologically produced bifunctional recombinant protein nanoparticles for immunoassays. / Jääskelainen, Anu; Harinen, Reija-Riitta; Soukka, Tero; Lamminmäki, Urpo; Korpimäki, Teemu; Virta, Marko.

In: Analytical Chemistry, Vol. 80, No. 3, 2008, p. 583-587.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Biologically produced bifunctional recombinant protein nanoparticles for immunoassays

AU - Jääskelainen, Anu

AU - Harinen, Reija-Riitta

AU - Soukka, Tero

AU - Lamminmäki, Urpo

AU - Korpimäki, Teemu

AU - Virta, Marko

PY - 2008

Y1 - 2008

N2 - 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. Ibis method of producing nanoparticles with inherent antigen binding activity presents several possibilities for the simple production of specific, functional nanoparticles. Production is fast, economical,and environmentally sustainable, making the system advantageous, particularly in applications requiring large quantities of specific nanoparticles.

AB - 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. Ibis method of producing nanoparticles with inherent antigen binding activity presents several possibilities for the simple production of specific, functional nanoparticles. Production is fast, economical,and environmentally sustainable, making the system advantageous, particularly in applications requiring large quantities of specific nanoparticles.

KW - 1182 Biochemistry, cell and molecular biology

U2 - 10.1021/ac071382v

DO - 10.1021/ac071382v

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VL - 80

SP - 583

EP - 587

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 3

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