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Diagnostic Mr 31 000 Schistosoma mansoni proteins: requirement of infection, but not immunization, and use of the “miniblot” technique for the production of monoclonal antibodies

Published online by Cambridge University Press:  05 June 2009

Andreas Ruppel ,
Ute Breternitz  and
Reinhard Burger
Andreas Ruppel
Affiliation:
Institut für Tropenhygiene, Im Neuenheimer Feld 324, D–6900 Heidelberg, Federal Republic of Germany
Ute Breternitz
Affiliation:
Institut für Immunologie, Universitüt Heidelberg, Federal Republic of Germany
Reinhard Burger
Affiliation:
Institut für Immunologie, Universitüt Heidelberg, Federal Republic of Germany
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Abstract

Antibodies directed against diagnostic Mr 31 000 polypeptide(s) of adult Schistosoma mansoni were already formed in mice during prepatency. In contrast, repeated immunization of mice with homogenates of adult schistosomes failed to elicit antibodies detectable in immunoblots in the Mr 31 000 region. Therefore, spleen cells of infected mice were used to produce hybridoma lines. The “miniblot technique” was developed in order to detect in hybridoma supernatants antibodies against schistosome Mr 31 000 components. Electrophoretically separated total S. mansoni proteins were transferred onto nitrocellulose, and the position of the Mr 31 000 components was determined with polyclonal antisera and immunoblotting. Pieces of about 3 square mm of nitrocellulose bearing the diagnostic proteins were incubated with about 100 μ1 of hybridoma supernatant in microtitre plates and subsequently probed with peroxidase-conjugated antibody to mouse IgG. This screening technique identified hybridomas secreting antibody to the relevant S. mansoni antigens. It is applicable to other defined parasit antigens, which are, however, not available in biochemically purified form. The monoclonal antibodies produced against the proteins with diagnostic potential reacted with antigens localized in the schistosome gut.

Type
Research Papers
Information
Journal of Helminthology , Volume 61 , Issue 2 , June 1987 , pp. 95 - 101
Copyright
Copyright © Cambridge University Press 1987

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