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Monoclonal Antibody Against Premembrane Viral Protein of Avian Tembusu Virus
Xiaozhi Huang1, Tingting Shi1, Kaikun Mo1
1Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University , Hangzhou, China .
Abstract:
Premembrane (prM) is a viral protein of flavivirus, which is important for the generation of infectious virion and for virus infection to the host. However, the biological properties and function of the prM of Avian Tembusu virus (ATMUV) have scarcely been studied to date. Monoclonal antibodies (mAbs) are a powerful tool for functional analysis of viral protein. To produce a mAb against prM protein of ATMUV, the prM gene sequence was amplified by reverse transcription polymerase chain reaction (RT-PCR) and cloned into the prokaryotic expression vector pET-28a (+). The recombinant prM protein was successfully expressed in BL21 (DE3). Using the purified prM as immunogen in mice, three hybridoma cells secreting mAbs against prM protein were obtained. These mAbs showed a strong reaction with ATMUV-infected DF-1 cells and pEGFP-C3-prM transfected 293-T cells in both Western blotting analysis and immunofluorescence assay. The mAbs developed in this study will be useful tools for analysis of the prM protein functions on ATMUV infection and the interaction between prM and its host molecules.
Insights
Researchers developed monoclonal antibodies (mAbs) against the Avian Tembusu virus (ATMUV) prM protein. These novel mAbs are valuable tools for studying ATMUV infection mechanisms and viral protein interactions.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- The premembrane (prM) protein is crucial for flavivirus infectivity and virion assembly.
- The specific functions of Avian Tembusu virus (ATMUV) prM protein remain largely uncharacterized.
- Monoclonal antibodies (mAbs) are essential for dissecting viral protein functions.
Purpose of the Study:
- To generate monoclonal antibodies (mAbs) targeting the prM protein of Avian Tembusu virus (ATMUV).
- To utilize these mAbs for analyzing the biological properties and functions of ATMUV prM.
Main Methods:
- Amplification and cloning of the ATMUV prM gene using RT-PCR and pET-28a (+) vector.
- Expression and purification of recombinant ATMUV prM protein in BL21 (DE3) cells.
- Production of hybridoma cell lines secreting mAbs against ATMUV prM via mouse immunization.
Main Results:
- Successfully produced three hybridoma cell lines secreting specific mAbs against ATMUV prM.
- Demonstrated strong reactivity of the generated mAbs with ATMUV-infected cells and prM-transfected cells.
- Validated mAb efficacy through Western blotting and immunofluorescence assays.
Conclusions:
- Developed novel mAbs specific to the ATMUV prM protein.
- These mAbs serve as effective tools for studying ATMUV infection dynamics.
- The antibodies will aid in understanding prM protein functions and host interactions during ATMUV infection.