Immunoblot analysis of natural and vaccine-induced IgG responses to rubella virus proteins expressed in insect cells

Jasminka Nedeljkovic, Tanjaq Jovanovic, Srecko Mladjenovic, Klaus Hedman, Nina Peitsaro, Christian Oker-Blom (Corresponding Author)

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18 Citations (Scopus)

Abstract

Background: The three structural proteins of rubella virus (RV), the capsid protein C and the envelope glycoproteins E1 and E2, were produced individually in soluble form in Sf9 insect cells using the baculovirus system. All proteins were equipped with a polyhistidine tag at their C-terminal ends to enable gentle purification by metal ion affinity chromatography. In addition, the E1 and E2 proteins were engineered to display the FLAG epitope tag at their N-terminal ends. Study design: The diagnostic potential of the recombinant purified proteins was evaluated by immunoblot and enzyme immuno assays (EIA) using a total of 57 well-characterised serum samples obtained at various time points after natural RV infection, congenital rubella syndrome (CRS), MMR vaccination or from controls with past RV immunity. In addition, acute and convalescent phase serum pools from a total of 20 patients were evaluated. Authentic RV proteins were used as a reference. Results: The recombinant E1 and C proteins were predominant in eliciting the immune responce in both postnatal and vaccinal RV infections, being much weaker in the vaccinal ones. The IgG responce to the recombinant C protein was very strong after the first month post infection and decreased with time. The immune responce against the recombinant E2 protein, however, was generally poor, but notably stronger after congenital infection. Together, the results showed that the individual recombinant protein antigens could be suitable for diagnosis of RV infection and for study of the immune response to rubella vaccination.
Original languageEnglish
Pages (from-to)119-131
Number of pages13
JournalJournal of Clinical Virology
Volume14
Issue number2
DOIs
Publication statusPublished - 1999
MoE publication typeA1 Journal article-refereed

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Rubella virus
Insects
Recombinant Proteins
Vaccines
Immunoglobulin G
Virus Diseases
Proteins
Vaccination
Congenital Rubella Syndrome
Sf9 Cells
Rubella
Baculoviridae
Enzyme Assays
Capsid Proteins
Protein C
Infection
Serum
Affinity Chromatography
Epitopes
Immunity

Cite this

Nedeljkovic, Jasminka ; Jovanovic, Tanjaq ; Mladjenovic, Srecko ; Hedman, Klaus ; Peitsaro, Nina ; Oker-Blom, Christian. / Immunoblot analysis of natural and vaccine-induced IgG responses to rubella virus proteins expressed in insect cells. In: Journal of Clinical Virology. 1999 ; Vol. 14, No. 2. pp. 119-131.
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abstract = "Background: The three structural proteins of rubella virus (RV), the capsid protein C and the envelope glycoproteins E1 and E2, were produced individually in soluble form in Sf9 insect cells using the baculovirus system. All proteins were equipped with a polyhistidine tag at their C-terminal ends to enable gentle purification by metal ion affinity chromatography. In addition, the E1 and E2 proteins were engineered to display the FLAG epitope tag at their N-terminal ends. Study design: The diagnostic potential of the recombinant purified proteins was evaluated by immunoblot and enzyme immuno assays (EIA) using a total of 57 well-characterised serum samples obtained at various time points after natural RV infection, congenital rubella syndrome (CRS), MMR vaccination or from controls with past RV immunity. In addition, acute and convalescent phase serum pools from a total of 20 patients were evaluated. Authentic RV proteins were used as a reference. Results: The recombinant E1 and C proteins were predominant in eliciting the immune responce in both postnatal and vaccinal RV infections, being much weaker in the vaccinal ones. The IgG responce to the recombinant C protein was very strong after the first month post infection and decreased with time. The immune responce against the recombinant E2 protein, however, was generally poor, but notably stronger after congenital infection. Together, the results showed that the individual recombinant protein antigens could be suitable for diagnosis of RV infection and for study of the immune response to rubella vaccination.",
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Immunoblot analysis of natural and vaccine-induced IgG responses to rubella virus proteins expressed in insect cells. / Nedeljkovic, Jasminka; Jovanovic, Tanjaq; Mladjenovic, Srecko; Hedman, Klaus; Peitsaro, Nina; Oker-Blom, Christian (Corresponding Author).

In: Journal of Clinical Virology, Vol. 14, No. 2, 1999, p. 119-131.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Immunoblot analysis of natural and vaccine-induced IgG responses to rubella virus proteins expressed in insect cells

AU - Nedeljkovic, Jasminka

AU - Jovanovic, Tanjaq

AU - Mladjenovic, Srecko

AU - Hedman, Klaus

AU - Peitsaro, Nina

AU - Oker-Blom, Christian

PY - 1999

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N2 - Background: The three structural proteins of rubella virus (RV), the capsid protein C and the envelope glycoproteins E1 and E2, were produced individually in soluble form in Sf9 insect cells using the baculovirus system. All proteins were equipped with a polyhistidine tag at their C-terminal ends to enable gentle purification by metal ion affinity chromatography. In addition, the E1 and E2 proteins were engineered to display the FLAG epitope tag at their N-terminal ends. Study design: The diagnostic potential of the recombinant purified proteins was evaluated by immunoblot and enzyme immuno assays (EIA) using a total of 57 well-characterised serum samples obtained at various time points after natural RV infection, congenital rubella syndrome (CRS), MMR vaccination or from controls with past RV immunity. In addition, acute and convalescent phase serum pools from a total of 20 patients were evaluated. Authentic RV proteins were used as a reference. Results: The recombinant E1 and C proteins were predominant in eliciting the immune responce in both postnatal and vaccinal RV infections, being much weaker in the vaccinal ones. The IgG responce to the recombinant C protein was very strong after the first month post infection and decreased with time. The immune responce against the recombinant E2 protein, however, was generally poor, but notably stronger after congenital infection. Together, the results showed that the individual recombinant protein antigens could be suitable for diagnosis of RV infection and for study of the immune response to rubella vaccination.

AB - Background: The three structural proteins of rubella virus (RV), the capsid protein C and the envelope glycoproteins E1 and E2, were produced individually in soluble form in Sf9 insect cells using the baculovirus system. All proteins were equipped with a polyhistidine tag at their C-terminal ends to enable gentle purification by metal ion affinity chromatography. In addition, the E1 and E2 proteins were engineered to display the FLAG epitope tag at their N-terminal ends. Study design: The diagnostic potential of the recombinant purified proteins was evaluated by immunoblot and enzyme immuno assays (EIA) using a total of 57 well-characterised serum samples obtained at various time points after natural RV infection, congenital rubella syndrome (CRS), MMR vaccination or from controls with past RV immunity. In addition, acute and convalescent phase serum pools from a total of 20 patients were evaluated. Authentic RV proteins were used as a reference. Results: The recombinant E1 and C proteins were predominant in eliciting the immune responce in both postnatal and vaccinal RV infections, being much weaker in the vaccinal ones. The IgG responce to the recombinant C protein was very strong after the first month post infection and decreased with time. The immune responce against the recombinant E2 protein, however, was generally poor, but notably stronger after congenital infection. Together, the results showed that the individual recombinant protein antigens could be suitable for diagnosis of RV infection and for study of the immune response to rubella vaccination.

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