Development of neutralizing antibody against the membrane glycoproteins of emerging disease viruses.
Date Issued
2016
Date
2016
Author(s)
Li, Chih-Chi
Abstract
The Middle East respiratory syndrome coronavirus (MERS-CoV) was first reported to infect human beings in September 2012. This virus seemed to develop into a highly contagious disease and caused high mortality rate up to 36%. Since December 2013 west Africa experienced the largest outbreak of Ebola, which was first discovered in 1976. The mortality of Ebola virus disease is relatively high, and yet the therapeutic drugs and vaccines are still under development. Researches for further understanding of the disease progression as well as accurate detection methods for these viruses are important for endemic control. Thus, the goal of this study is to develop specific antibodies against Ebola virus or MERS-CoV for the application in virus detection and clinical therapy. Two of the virus surface proteins, glycoprotein（GP）and spike protein（SP）, which were predicted as having high specificity and antigenicity were chosen as the antigens. In Ebola case, 3 peptides fragments of GP and its truncated form, constructed in refer to the published anti-GP mAb KZ52, were synthesized and expressed for mice immunization. In MERS-CoV case, S1 domain and receptor binding domain（RBD）were constructed and expressed in Bac-to-Bac Expression system for immunization. After repeat boosts, mice were sacrificed for producing hybridoma. Monoclonal antibodies（mAb） were screened by enzyme-linked immunosorbent assay（ELISA）and then identified by Western blot. Finally, neutralizing assays for MERS-CoV were used to verify the inhibition of the specific mAb which might block the virus invasion through the expressed DPP4 receptor on the cell surface. Useful mAb will be selected and used to develop the blocking ELISA or the antigen-capture ELISA, which could be utilized in clinical diagnosis or quick viral detection in the field.
Subjects
Ebola virus
MERS-CoV
Glycoprotein
Spike protein
neutralizing monoclonal antibody
SDGs
Type
thesis
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