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Partial characterization of 2-β-mercaptoethanol-soluble surface-associated antigens of Onchocerca volvulus

Published online by Cambridge University Press:  06 April 2009

F. Engelbrecht
Affiliation:
Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 IQP, UK
G. Braun
Affiliation:
Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 IQP, UK
V. Connor
Affiliation:
Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 IQP, UK
M. Downham
Affiliation:
MRC Laboratory, PO Box 81, Bo, Sierra Leone
J. A. Whitworth
Affiliation:
MRC Laboratory, PO Box 81, Bo, Sierra Leone
D. W. Taylor
Affiliation:
Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 IQP, UK

Extract

Antigens were extracted from the epicuticle/cuticle of intact female Onchocerca volvulus using 2% 2-β-mercaptoethanol and 1% SDS. In Western blot analysis a human infection serum selected for its high antibody titre against whole worm homogenates did not recognize any component solubilized by 1% SDS. However, the same serum did bind at least 7 antigens among the material extracted with 2-β-mercaptoethanol. These antigens have apparent molecular weights (Mr) of: 15 000, 18 000, 28 000, 78 000, 98 000, 120 000 and 200 000. In ELISA using this preparation as target antigen, 151 out of 153 human infection sera gave positive results. An Onchocerca-specific IgG1 monoclonal antibody, designated Cam1, recognized the 28 000 Mr antigen, which is the most prominent antigen detected by Western blot analysis using human infection sera. In ELISA, using material affinity-purified with Cam1 as target antigen, 149 out of 153 human infection sera gave a positive IgG response. From a cDNA library three expressing clones were isolated with a rabbit serum raised against 2-β-mercaptoethanol solubilized material. One of these clones was recognized by the monoclonal antibody Cam1.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1991

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