The use of dextran as a blocking agent on nitrocellulose membrane in the analysis of sporozoite antigens of Plasmodium vivax.
Dextran (molecular weight, 71,200) has been found to block the unbound sites of the nitrocellulose membrane, to which antigens have been electroblotted from acrylamide gel, for use in assaying monoclonal antibodies. The use of polysaccharide as a blocking agent allows the antigens on the nitrocellulose membrane to be digested with pronase and subsequently reacted with monoclonal antibodies. Sporozoite antigens of Plasmodium vivax, after being digested with pronase, completely lost their antigenicity to bind to the sporozoite-specific monoclonal antibodies, thus suggesting that they are proteins or protein conjugates in nature. The method described here for qualitative determination of protein antigens requires as little as 2000 sporozoites for each assay.
Yuan
L F
LF
Naval Medical Research Institute, Bethesda, MD 20014.
Wirtz
R A
RA
Beaudoin
R L
RL
D000818:Animals; D000913:Antibodies, Protozoan / Q000032:analysis*; D000953:Antigens, Protozoan / Q000032:analysis*; D001665:Binding Sites / Q000187:drug effects; D015153:Blotting, Western / Q000379:methods; D003101:Collodion; D003911:Dextrans / Q000494:pharmacology; D010966:Plasmodium vivax / Q000276:immunology; D011402:Pronase
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D003911:Dextrans; D003101:Collodion; D011402:Pronase