Characterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid ProteinReport as inadecuate




Characterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid Protein - Download this document for free, or read online. Document in PDF available to download.

The transmissible gastroenteritis virus TGEV nucleocapsid N protein plays important roles in the replication and translation of viral RNA. The present study provides the first description of two monoclonal antibodies mAbs 5E8 and 3D7 directed against the TGEV N protein linker region LKR and carboxyl terminal domain CTD. The mAbs 5E8 and 3D7 reacted with native N protein in western blotting and immunofluorescence assay IFA. Two linear epitopes, 189SVEQAVLAALKKLG202 and 246VTRFYGARSSSA257, located in the LKR and CTD of TGEV N protein, respectively, were identified after truncating the protein and applying a peptide scanning technique. Using mAb 5E8, we observed that the N protein was expressed in the cytoplasm during TGEV replication and that the protein could be immunoprecipitated from TGEV-infected PK-15 cells. The mAb 5E8 can be applied for different approaches to diagnosis of TGEV infection. In addition, the antibodies represent useful tools for investigating the antigenic properties of the N protein.



Author: Xin Zhang , Xin Zhao , Hui Dong , Yunnuan Zhu, Hongyan Shi, Jianfei Chen, Da Shi, Li Feng

Source: http://plos.srce.hr/



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