Abstract
This study describes the identification of one linear B-cell epitope on TMUV NS1 protein with monoclonal antibody (mAb) 3G2 by indirect enzyme-linked immunosorbent assay (ELISA). In this study, NS1 protein was expressed in prokaryotic expression system and purified. One mAb against NS1 protein was generated from Balb/c mice immunized with recombinant protein NS1. A set of 35 partially-overlapping polypeptides covering the entire NS1 protein was expressed with PGEX-6P-1 vector and screened with mAb 3G2. One polypeptide against the mAb was acquired and identified by indirect ELISA and western-blot. To map the epitope accurately, one or two amino acid residues were removed from the carboxy and amino terminal of polypeptide sequentially. A series of truncated oligopeptides were expressed and purified. The minimal determinant of the linear B cell epitope was recognized and identified with mAb 3G2. The accurate linear B-cell epitope was 269DEKEIV274 located in NS1 protein. Furthermore, sequence alignment showed that the epitope was highly conserved and specific among TMUV strains and other flavivirus respectively. The linear B-cell epitope of TMUV NS1 protein could benefit the development of new vaccines and diagnostic assays.
Highlights
Tembusu virus infection in ducks is caused by Tembusu virus (TMUV), which was first reported in April 2010 in China[1]
NS1 protein is the non-structural protein of flavivirus, which has important functional properties in virus RNA replication, anti-viral immunity and host recognition of virus-associated molecular patterns[11, 21,22,23]
It is reported that NS1 protein or monoclonal antibody (mAb) against NS1 protein can confer active or passive immunity protection from the flavivirus challenge [24,25,26], which is closely related to the epitopes on NS1 protein
Summary
Tembusu virus infection in ducks is caused by Tembusu virus (TMUV), which was first reported in April 2010 in China[1]. The virus can infect more varieties of ducks such as Beijing duck, golden duck, Shaoxing duck, Cherry Valley, Campbell ducks, Jinyun duck, etc. Laying ducks infected mainly demonstrated drops in egg production, follicular rupture and bleeding, and yolk peritonitis. Ducklings mainly displayed standing instability and paralysis, retarded growth, with 10 to 30% mortality rates [2, 3]. Other birds such as Chickens, geese, sparrows, etc were infected and displayed obvious clinical signs [4,5,6].
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