Production and characterization of virus-like particles and the P domain protein of GII.4 norovirus

• Published in: Journal of virological methods Vol. 179; no. 1; pp. 1 - 7
• Main Authors: Koho, Tiia, Huhti, Leena, Blazevic, Vesna, Nurminen, Kirsi, Butcher, Sarah J., Laurinmäki, Pasi, Kalkkinen, Nisse, Rönnholm, Gunilla, Vesikari, Timo, Hytönen, Vesa P., Kulomaa, Markku S.
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01.01.2012; Elsevier
• Subjects: Animals; antibodies; Antibodies, Viral - blood; Biological and medical sciences; Caliciviridae Infections - diagnosis; Caliciviridae Infections - prevention & control; capsid; Cell Line; cryo-electron microscopy; Diagnostics; enzyme-linked immunosorbent assay; Escherichia coli; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; gastroenteritis; Gene Expression; Humans; Immunoassay; insects; light scattering; Microbiology; Norovirus; Norovirus - genetics; Norovirus - immunology; P domain protein; proteins; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Spodoptera; Techniques used in virology; vaccination; Viral Proteins - genetics; Viral Proteins - immunology; Viral Vaccines - immunology; Virology; Virosomes - genetics; Virosomes - immunology; Virosomes - isolation & purification; Virosomes - metabolism; Virus-like particle; virus-like particles; VLP; Virus-like particle; Norovirus; VLP; Diagnostics; P domain protein; Virus; Enzyme; Caliciviridae; Virus like particle; Oxidoreductases; Method; Glycine dehydrogenase (decarboxylating)
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2011.05.009

Noroviruses are an important cause of epidemic acute gastroenteritis in humans. In this study the production and characterization of GII.4 norovirus virus-like particles (VLPs) in insect cells is reported. Furthermore, the expression of corresponding norovirus polyhistidine-tagged P domain protein in Escherichia coli is described. The protruding P domain of the norovirus capsid is known to contain determinants for antibody and receptor binding. Therefore, P domain proteins were studied as an alternative diagnostic tool for evaluating norovirus infection. Analyses by dynamic light scattering and cryo-electron microscopy revealed the presence of intact VLPs with an average diameter of about 40nm. Immunostaining and ELISA assays using norovirus-specific human sera revealed that VLPs and the P domain are recognized by norovirus-specific antibodies and by their putative receptor. The VLPs and P domain protein are potentially useful in the development of diagnostic and vaccination tools for noroviruses.