Immunological reactivity of baculovirus-expressed enterovirus proteins

L. Dahllund (Corresponding Author), Christian Oker-Blom, Michel Schmidt, T. Hyypiä

Research output: Contribution to journalArticleScientificpeer-review

Abstract

In order to study immunological reactivity of individual enterovirus polypeptides and evaluate their usefulness for enterovirus diagnosis, the genes coding for viral structural and nonstructural proteins were expressed using recombinant baculoviruses. A histidine-tailed coxsackievirus B3 (CBV3) VP1 capsid protein was expressed and purified by immobilized metal ion affinity chromatography for EIAs. To elucidate the usefulness of the other CBV3 capsid proteins for immunoassays, recombinant baculovirus expressing the whole CBV3 capsid polyprotein region was constructed. For the detection of a potentially broader spectrum of enteroviruses, the conserved nonstructural P3 region was expressed. The P3 region encodes four nonstructural proteins including a cysteine protease (3C) and an RNA-dependent RNA-polymerase (3D). The 3C polypeptide was shown to be proteolytically active indicating that the baculovirus system is capable of expressing biologically functional enterovirus proteins. Immunoblot analysis detected antibodies against the VP1, 3C and 3D proteins in human serum samples. When the baculovirus-expressed antigens were compared with lysates of enterovirus-infected cells and a synthetic peptide in EIA highly similar results were obtained with recombinant VP1 and the lysate antigens. Although reactive in immunoblots, the P3 encoded proteins were not satisfactory for EIA.

Original languageEnglish
Pages (from-to)209 - 220
Number of pages12
JournalJournal of Virological Methods
Volume67
Issue number2
DOIs
Publication statusPublished - 1997
MoE publication typeA1 Journal article-refereed

Fingerprint

Enterovirus
Baculoviridae
Proteins
Capsid Proteins
Peptides
Viral Nonstructural Proteins
Artificial Cells
Viral Structural Proteins
RNA Replicase
Antigens
Polyproteins
Cysteine Proteases
Capsid
Affinity Chromatography
Immunoassay
Histidine
Metals
Ions
Antibodies
Serum

Cite this

Dahllund, L. ; Oker-Blom, Christian ; Schmidt, Michel ; Hyypiä, T. / Immunological reactivity of baculovirus-expressed enterovirus proteins. In: Journal of Virological Methods. 1997 ; Vol. 67, No. 2. pp. 209 - 220.
@article{5f27c2b29a74416fa354481b6c35fd32,
title = "Immunological reactivity of baculovirus-expressed enterovirus proteins",
abstract = "In order to study immunological reactivity of individual enterovirus polypeptides and evaluate their usefulness for enterovirus diagnosis, the genes coding for viral structural and nonstructural proteins were expressed using recombinant baculoviruses. A histidine-tailed coxsackievirus B3 (CBV3) VP1 capsid protein was expressed and purified by immobilized metal ion affinity chromatography for EIAs. To elucidate the usefulness of the other CBV3 capsid proteins for immunoassays, recombinant baculovirus expressing the whole CBV3 capsid polyprotein region was constructed. For the detection of a potentially broader spectrum of enteroviruses, the conserved nonstructural P3 region was expressed. The P3 region encodes four nonstructural proteins including a cysteine protease (3C) and an RNA-dependent RNA-polymerase (3D). The 3C polypeptide was shown to be proteolytically active indicating that the baculovirus system is capable of expressing biologically functional enterovirus proteins. Immunoblot analysis detected antibodies against the VP1, 3C and 3D proteins in human serum samples. When the baculovirus-expressed antigens were compared with lysates of enterovirus-infected cells and a synthetic peptide in EIA highly similar results were obtained with recombinant VP1 and the lysate antigens. Although reactive in immunoblots, the P3 encoded proteins were not satisfactory for EIA.",
author = "L. Dahllund and Christian Oker-Blom and Michel Schmidt and T. Hyypi{\"a}",
year = "1997",
doi = "10.1016/S0166-0934(97)00098-0",
language = "English",
volume = "67",
pages = "209 -- 220",
journal = "Journal of Virological Methods",
issn = "0166-0934",
publisher = "Elsevier",
number = "2",

}

Immunological reactivity of baculovirus-expressed enterovirus proteins. / Dahllund, L. (Corresponding Author); Oker-Blom, Christian; Schmidt, Michel; Hyypiä, T.

In: Journal of Virological Methods, Vol. 67, No. 2, 1997, p. 209 - 220.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Immunological reactivity of baculovirus-expressed enterovirus proteins

AU - Dahllund, L.

AU - Oker-Blom, Christian

AU - Schmidt, Michel

AU - Hyypiä, T.

PY - 1997

Y1 - 1997

N2 - In order to study immunological reactivity of individual enterovirus polypeptides and evaluate their usefulness for enterovirus diagnosis, the genes coding for viral structural and nonstructural proteins were expressed using recombinant baculoviruses. A histidine-tailed coxsackievirus B3 (CBV3) VP1 capsid protein was expressed and purified by immobilized metal ion affinity chromatography for EIAs. To elucidate the usefulness of the other CBV3 capsid proteins for immunoassays, recombinant baculovirus expressing the whole CBV3 capsid polyprotein region was constructed. For the detection of a potentially broader spectrum of enteroviruses, the conserved nonstructural P3 region was expressed. The P3 region encodes four nonstructural proteins including a cysteine protease (3C) and an RNA-dependent RNA-polymerase (3D). The 3C polypeptide was shown to be proteolytically active indicating that the baculovirus system is capable of expressing biologically functional enterovirus proteins. Immunoblot analysis detected antibodies against the VP1, 3C and 3D proteins in human serum samples. When the baculovirus-expressed antigens were compared with lysates of enterovirus-infected cells and a synthetic peptide in EIA highly similar results were obtained with recombinant VP1 and the lysate antigens. Although reactive in immunoblots, the P3 encoded proteins were not satisfactory for EIA.

AB - In order to study immunological reactivity of individual enterovirus polypeptides and evaluate their usefulness for enterovirus diagnosis, the genes coding for viral structural and nonstructural proteins were expressed using recombinant baculoviruses. A histidine-tailed coxsackievirus B3 (CBV3) VP1 capsid protein was expressed and purified by immobilized metal ion affinity chromatography for EIAs. To elucidate the usefulness of the other CBV3 capsid proteins for immunoassays, recombinant baculovirus expressing the whole CBV3 capsid polyprotein region was constructed. For the detection of a potentially broader spectrum of enteroviruses, the conserved nonstructural P3 region was expressed. The P3 region encodes four nonstructural proteins including a cysteine protease (3C) and an RNA-dependent RNA-polymerase (3D). The 3C polypeptide was shown to be proteolytically active indicating that the baculovirus system is capable of expressing biologically functional enterovirus proteins. Immunoblot analysis detected antibodies against the VP1, 3C and 3D proteins in human serum samples. When the baculovirus-expressed antigens were compared with lysates of enterovirus-infected cells and a synthetic peptide in EIA highly similar results were obtained with recombinant VP1 and the lysate antigens. Although reactive in immunoblots, the P3 encoded proteins were not satisfactory for EIA.

U2 - 10.1016/S0166-0934(97)00098-0

DO - 10.1016/S0166-0934(97)00098-0

M3 - Article

VL - 67

SP - 209

EP - 220

JO - Journal of Virological Methods

JF - Journal of Virological Methods

SN - 0166-0934

IS - 2

ER -