Mimicking rubella virus particles by using recombinant envelope glycoproteins and liposomes.
The envelope glycoproteins E1 and E2 of rubella virus (RV) were engineered to display the FLAG epitope tag and a polyhistidine tag, at their amino and carboxy termini, respectively. These modified envelope proteins were produced in Sf9 insect cells utilizing baculovirus expression vectors, the E1 and E2 vectors giving rise to protein products of about 58 and 42 kDa, respectively. The recombinant proteins were purified by immobilized metal-ion affinity chromatography and reconstituted into liposomes via their hydrophobic transmembrane anchors. The liposomes were prepared by detergent dialysis in the presence of europium-DTPA chelate, enabling the subsequent measurement of the binding of the resultant proteoliposomes to the antibodies by time resolved fluorescence. RV mimicking proteoliposomes were recognized by antibodies specific for the E1 and E2 proteins, as well as the FLAG epitope tag. This type of virosome may prove useful for studies on the basic biological events of an RV infection or as diagnostic reagents.
Orellana
A
A
VTT Biotechnology and Food Research, Finland.
Mottershead
D
D
van der Linden
I
I
Keinänen
K
K
Oker-Blom
C
C
D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000235:genetics; D002614:Chelating Agents / Q000737:chemistry; D005063:Europium / Q000737:chemistry; D006023:Glycoproteins / Q000235:genetics; D015151:Immunoblotting; D007915:Lepidoptera / Q000166:cytology / Q000235:genetics; D008081:Liposomes / Q000737:chemistry*; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D004369:Pentetic Acid / Q000737:chemistry; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012409:Rubella / Q000175:diagnosis; D012412:Rubella virus / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
D000956:Antigens, Viral; D002614:Chelating Agents; D006023:Glycoproteins; D008081:Liposomes; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C061826:E1 envelope protein, Rubella virus; C061827:E2 envelope protein, Rubella virus; D005063:Europium; D004369:Pentetic Acid