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Immunological detection of Plasmodium knowlesi antigens after electrophoretic transfer from SDS‐polyacrylamide gels to diazophenylthioether paper
Author(s) -
JOHNSON DIANA M. A.,
COHEN S.
Publication year - 1984
Publication title -
parasite immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.795
H-Index - 75
eISSN - 1365-3024
pISSN - 0141-9838
DOI - 10.1111/j.1365-3024.1984.tb00783.x
Subject(s) - plasmodium knowlesi , antigenicity , polyclonal antibodies , antigen , biology , polyacrylamide gel electrophoresis , epitope , antibody , monoclonal antibody , gel electrophoresis , microbiology and biotechnology , antiserum , virology , plasmodium falciparum , biochemistry , immunology , malaria , enzyme , plasmodium vivax
Summary Freeze‐thawed extracts of mature schizonts of Plasmodium knowlesi (strain Wl) were electrophoresed in sodium dodecylsulphate (SDS) polyacryl‐amide gels, then transferred electrophoretically to diazophenylthioether paper. The transferred antigen was probed using a purified polyvalent, polyclonal immune serum pool isolated from rhesus monkeys ( Macaca mulatto ). This method for identifying parasite antigens was compared with immunoabsorption, using parasites which had been metabolically labelled in vitro with 35 S‐methionine. The methods showed general correspondence but some antigens failed to react after transfer because of loss of antigenicity during SDS‐polyacramide gel electrophoresis (SDS‐PAGE). Some antigens were observed preferentially after the transfer procedure, possibly because of their failure to elute from the immunoabsorbant or because they may have a low methionine content. The transfer method failed to reveal the antigen reactivity of three out of four monoclonal antibodies tested, apparently because the epitopes involved were inactivated by SDS. During P. knowlesi infection in the resistant kra monkey ( Macaca fascicularis ) the transfer method showed the presence by day 10 of antibodies against two components having an apparent molecular weight by SDS‐PAGE similar to those of putative protective antigens described in other studies using monoclonal antibodies.