To study the antigenic characteristics and differentiation serotypes of Hantavirus, the produced hybridoma secreting monoclonal antibodies(MAbs) against Hantaan virus, 76/118 strain and Seoul virus, 80/39 strain were produced by fusion of mouse
myeloma
cell,
Sp2/0-Ag14 with spleen cells isolated from Balb/c mice immunized with Hantaan virus or Seoul virus. From the results in this study, the MAbs against Hantaan virus were screened by indirect immunofluorescent antibody techinque(IFA). 587 hybridoma
cells
were produced out of 768 wells and the fusion frequency was 76%. Seven among 587 hybridoma cells produced MAbs against Hantaan virus continuously. Isotype analysis of MAbs against Hantaan virus revealed that all these MAbs belong to the IgG2a
subclass.
All 7 MAbs reacted to nucleocapsid(N)proteins(MW.49-50Kd) of Hantaan virus by immunoblot assay.
MAbs against Seoul virus were screened, 985 hybridoma cells were produced out of 1152 and the fusion frequency was 86%. Eight among 985hybridoma cells produced MAbs against Seoul virus continuously. Isotype analysis of these MAbs against Seoul
virus
revealed the IgM and IgG2a subelass. These 8 MAbs reacted to nucleocapsid (N) proteins (MW.49-50Kd) of Seoul virus as demonstrated by immunoblot. Interestingly, these MAbs had neutralizing activity deter mined by PRNT analysis. Therefore it is
suspected
that the antigenic sites on N protein
may be involved in the virus neutralization.
The serological reactivity of 7 MAbs against Hantaan virus with Hantaan virus were strong, although Seoul, Puumala and prospect hill virus were not reacted to these MAbs. 8 MAbs against Seoul virus, reacted to Seoul virus strongly. Hantaan and
Puumala
virus reacted slightl to these MAbs but Prospect hill virus did not.
To comparative study of sensitivity test between IFA and double sandwich ELISA method for human sera using ELISA of which these MAbs coated were tested, the ELISA method was more sensitive than IFA.
These MAbs reacted specifically to Hantaan and Seoul virus. there are antigenic relationships among Hantaviruses could be analysed by using these MAbs.
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