An economical procedure for screening of hybridoma supernatants for surface reactive antibodies to filarial larvae.
Third- and fourth-stage larvae of Onchocerca volvulus can be attached to the lids of 96-well microtiter plates to facilitate the screening of hybridoma supernatants reactive with filarial larvae. Plates are coated with a solution consisting of 0.1% gelatin, and 0.01% chrome alum. Following coating of plate lids one larva per depression is placed on the lid. An immunofluorescence assay (IFA) was conducted in which there were two 45 minute incubations and eight separate washes by immersion in PBS. At the completion of the assay over 90% of the larvae remain attached. The background in IFA is minimal and the results are consistent with those obtained using a suspension IFA. This technique permits rapid identification of antibody producing hybridomas with small numbers of larvae.
Miller
K M
KM
Lindsley F. Kimball Research Institute, New Blood Center, New York, N.Y.
Hotze
C
C
Brotman
B
B
Prince
A M
AM
D000818:Animals; D000909:Antibodies, Helminth / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006825:Hybridomas / Q000276:immunology*; D007814:Larva / Q000276:immunology; D009854:Onchocerca / Q000276:immunology*
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal