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Volume 27, Number 12—December 2021
Research

Novel Assay to Measure Seroprevalence of Zika Virus in the Philippines

Cameron Adams, Ramesh Jadi, Bruno Segovia-Chumbez, Jedas Daag, Michelle Ylade, Freddy A. Medina, Tyler M. Sharp, Jorge L. Munoz-Jordan, In-Kyu Yoon, Jacqueline Deen, Anna Lena Lopez1, Aravinda M. de Silva, and Lakshmanane PremkumarComments to Author 
Author affiliations: University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA (C. Adams, R. Jadi, B. Segovia-Chumbez, A.M. de Silva, L. Premkumar); University of the Philippines, Manila, Philippines (J. Daag, M. Ylade, J. Deen, A.L. Lopez); Centers for Disease Control and Prevention, San Juan, Puerto Rico, USA (F.A. Medina, T.M. Sharp, J.L. Munoz-Jordan); Coalition for Epidemic Preparedness Innovations, Washington, DC, USA (I.-K. Yoon)

Main Article

Figure 1

ZIKV Biotinylated-EDIII antigen capture ELISA in study of novel assay to measure ZIKV seroprevalence in the Philippines. A) Schematic of the ZIKV EDIII construct with an N-terminal human albumin secretion signal, a HaloTag (Promega, https://www.promega.com) for site-specific biotinylation and a C-terminal 6-histidine residue tag for affinity purification. The EDIII construct design also included a TEV protease cleavage site between HaloTag and EDIII. B) Biotinylated ZIKV EDIII displays an electrophoretic mobility shift with streptavidin. A site-specific biotinylated ZIKV EDIII was prepared using HaloTag biotin ligand. Electrophoretic gel shift analysis was performed in sodium dodecyl sulphate–polyacrylamide gel electrophoresis with biotinylated EDIII antigen in the presence and absence of streptavidin. C) Schematic of second-generation ZIKV EDIII ELISA using streptavidin-biotin interaction. Biotinylated EDIII antigen is captured by plate immobilized streptavidin. The antibody bound to EDIII is detected by a secondary anti–human IgG conjugated to alkaline phosphatase. EDIII, E protein domain III; IgG-AP, secondary IgG antibody conjugated to alkaline phosphatase; SA, streptavidin; SP, N terminal human albumin secretion signal; TEV, tobacco etch virus protease cleavage site; ZIKV, Zika virus.

Figure 1. ZIKV Biotinylated-EDIII antigen capture ELISA in study of novel assay to measure ZIKV seroprevalence in the Philippines. A) Schematic of the ZIKV EDIII construct with an N-terminal human albumin secretion signal, a HaloTag (Promega, https://www.promega.com) for site-specific biotinylation and a C-terminal 6-histidine residue tag for affinity purification. The EDIII construct design also included a TEV protease cleavage site between HaloTag and EDIII. B) Biotinylated ZIKV EDIII displays an electrophoretic mobility shift with streptavidin. A site-specific biotinylated ZIKV EDIII was prepared using HaloTag biotin ligand. Electrophoretic gel shift analysis was performed in sodium dodecyl sulphate–polyacrylamide gel electrophoresis with biotinylated EDIII antigen in the presence and absence of streptavidin. C) Schematic of second-generation ZIKV EDIII ELISA using streptavidin-biotin interaction. Biotinylated EDIII antigen is captured by plate immobilized streptavidin. The antibody bound to EDIII is detected by a secondary anti–human IgG conjugated to alkaline phosphatase. EDIII, E protein domain III; IgG-AP, secondary IgG antibody conjugated to alkaline phosphatase; SA, streptavidin; SP, N terminal human albumin secretion signal; TEV, tobacco etch virus protease cleavage site; ZIKV, Zika virus.

Main Article

1Deceased.

Page created: August 25, 2021
Page updated: November 19, 2021
Page reviewed: November 19, 2021
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