Broad-spectrum monoclonal antibodies against chikungunya virus structural proteins: Promising candidates for antibody-based rapid diagnostic test development
Fig 4
Protein regions targeted by monoclonal antibodies (mAbs) raised against chikungunya virus (CHIKV).
(a) Indirect immunofluorescent test of mAbs from strategy B against cells transfected with plasmids encoding HA-tagged full-length or truncated versions of CHIKV (CP10 strain) E3-E2-6K-E1, 6K-E1, E1, or 6K envelope proteins. CK47 was used as anti-E1 control antibody. (b) mAbs from strategy C against cells transfected with plasmid encoding capsid protein of CHIKV (CP10 strain). 19B02 was used as an anti-C control antibody. Cells that reacted with the 2nd antibody only were used as a negative control. Luciferase counts within culture supernatants expressed from a co-transfected luciferase-secreting plasmid (pMetLuc-Control, Ready-To-Glow Secreted Luciferase Reporter Assay, Takara Bio USA, Inc.) are shown. The reactivity of each mAb was detected by Alexa Fluor 488-conjugated secondary antibody (green color); expression of the HA peptide was detected by Alexa Fluor 555-conjugated secondary antibody (orange/red color); DAPI DNA counterstain was used to stain nuclei of cells (blue color). Images are representative of results obtained from two independent experiments and were taken under 40x objective magnification using an Axio observer Z.1 microscope (Carl Zeiss). The multiple-color images were generated by merging using ImageJ 1.5Oi (National Institutes of Health USA).