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Biochemical and Biophysical Research Communications, 2019. 513(4), 781-786, DOI: https://doi.org/10.1016/j.bbrc.2019.03.212
Construction of the safe neutralizing assay system using pseudotyped Nipah virus and G protein-specific monoclonal antibody
Seong Eun Bae, Seong Su Kim;Seong Tae Moon;Yeon Dong Cho;Hansaem Lee;Joo-Yeon Lee;Ha Youn Shin;Hee-Jung Lee;Young Bong Kim
• Construction of pseudotyped Henipavirus based on MuLV system using TELCeB6 cell.
• Production of monoclonal antibodies specific Nipah virus G glycoprotein.
• Neutralizing assay of monoclonal antibodies using pseudotyped Henipavirus.
• Isolation of monoclonal antibodies with neutralizing activity.
Nipah virus (NiV) is a recently emerged paramyxovirus that causes acute respiratory illness and fatal encephalitis in a broad spectrum of vertebrates, including humans. Due to its high pathogenicity and mortality rates, NiV requires handling in biosafety level-4 (BSL-4) containment facilities and no effective vaccines or therapeutic agents are currently available. Since current diagnostic tests for detecting serum neutralizing antibodies against NiV mainly employ live viruses, establishment of more safe and robust alternative diagnostic methods is an essential medical requirement. Here, we have developed a pseudotyped NiV and closely related Hendra virus (HeV) expressing envelope attachment (G) and fusion (F) glycoproteins using the Moloney murine leukemia virus (MuLV) packaging system. We additionally generated polyclonal antibodies (pAbs) against NiV-G and HeV-G and assessed their neutralizing activities for potential utilization in the pseudovirus-based neutralization assay and further application in the serum diagnostic test. To enhance the specificity of neutralizing antibody and sensitivity of the serological diagnostic test, monoclonal antibodies (mAbs) against NiV-G were generated, and among which four out of six mAb clones showed significant reactivity. Specifically, the 7G9 clone displayed the highest sensitivity. The selected mAb clones showed no cross-reactivity with HeV-G and efficient neutralizing activities against pseudotyped NiV. These results validate the safety and specificity of neutralization assays against NiV and HeV and present a useful tool to design effective vaccines and serological diagnosis.
- DOI: https://doi.org/10.1016/j.bbrc.2019.03.212
- ISBN or ISSN: 0006-291X
- 본 연구는 질병관리본부 연구개발과제연구비를 지원받아 수행되었습니다.
- This research was supported by a fund by Research of Korea Centers for Disease Control and Prevention.