title: Expression and Purification of E2 Glycoprotein from Insect Cells (Sf9) for Use in Serology.
abstract: Chikungunya virus (CHIKV) is a mosquito-borne arbovirus which poses a major threat to global public health. Definitive CHIKV diagnosis is crucial, especially in distinguishing the disease from dengue virus, which co-circulates in endemic areas and shares the same mosquito vectors. Laboratory diagnosis is mainly based on serological or molecular approaches. The E2 glycoprotein is a good candidate for serological diagnosis since it is the immunodominant antigen during the course of infection, and reacts with seropositive CHIKV sera. In this chapter, we describe the generation of stable clone Sf9 (Spodoptera frugiperda) cells expressing secreted, soluble, and native recombinant CHIKV E2 glycoprotein. We use direct plasmid expression in insect cells, rather than the traditional technique of generating recombinant baculovirus. This recombinant protein is useful for serological diagnosis of CHIKV infection.
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, 50603, Malaysia.
authors.forename: Chong Long
authors.initials: CL
authors.lastname: Chua
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, 50603, Malaysia.
authors.forename: I-Ching
authors.initials: IC
authors.lastname: Sam
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, 50603, Malaysia. chanyf@ummc.edu.my.
authors.forename: Yoke Fun
authors.initials: YF
authors.lastname: Chan
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000378:metabolism*; D002646:Chikungunya virus / Q000276:immunology / Q000378:metabolism*; D016056:Immunodominant Epitopes / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D012698:Serologic Tests; D061987:Sf9 Cells; D014759:Viral Envelope Proteins / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D016056:Immunodominant Epitopes; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins
keywords: Chikungunya; E2 glycoprotein; Enzyme-linked immunosorbent assay; Immunodominant antigen; Recombinant protein; Serology; Sf9