Kubickova, B.B.KubickovaSchenk, J.A.J.A.SchenkRamm, F.F.RammMarkuskiene, K.K.MarkuskieneReetz, J.J.ReetzDremsek, P.P.DremsekTamosiunas, P.L.P.L.TamosiunasCepulyte, L.L.CepulyteTrinh, H.A.H.A.TrinhScholz, J.J.ScholzMemczak, H.H.MemczakHovestädt, M.M.HovestädtRyll, R.R.RyllPetraityte-Burneikiene, R.R.Petraityte-BurneikieneCorman, V.M.V.M.CormanAndersson, A.A.AnderssonBecher, D.D.BecherGroschup, M.H.M.H.GroschupKubick, S.S.KubickSellrie, F.F.SellrieJohne, R.R.JohneUlrich, R.G.R.G.Ulrich2022-03-062022-03-062021https://publica.fraunhofer.de/handle/publica/27043610.1007/s00253-021-11342-7To generate a hepatitis E virus (HEV) genotype 3 (HEV-3)-specific monoclonal antibody (mAb), the Escherichia coli-expressed carboxy-terminal part of its capsid protein was used to immunise BALB/c mice. The immunisation resulted in the induction of HEV-specific antibodies of high titre. The mAb G117-AA4 of IgG1 isotype was obtained showing a strong reactivity with the homologous E. coli, but also yeast-expressed capsid protein of HEV-3. The mAb strongly cross-reacted with ratHEV capsid protein derivatives produced in both expression systems and weaker with an E. coli-expressed batHEV capsid protein fragment. In addition, the mAb reacted with capsid protein derivatives of genotypes HEV-2 and HEV-4 and common vole hepatitis E virus (cvHEV), produced by the cell-free synthesis in Chinese hamster ovary (CHO) and Spodoptera frugiperda (Sf21) cell lysates. Western blot and line blot reactivity of the mAb with capsid protein derivatives of HEV-1 to HEV-4, cvHEV, ratHEV and batHEV suggested a linear epitope. Use of truncated derivatives of ratHEV capsid protein in ELISA, Western blot, and a Pepscan analysis allowed to map the epitope within a partially surface-exposed region with the amino acid sequence LYTSV. The mAb was also shown to bind to human patient-derived HEV-3 from infected cell culture and to hare HEV-3 and camel HEV-7 capsid proteins from transfected cells by immunofluorescence assay. The novel mAb may serve as a useful tool for further investigations on the pathogenesis of HEV infections and might be used for diagnostic purposes.en610660620journal article