| Abstract |
LOU/C rat as wells as BALB/c mice were immunized with male Xenopus plasma
from animals stimulated by 17-beta-oestradiol. Both species reacted equally
well after two i.p. inoculations of 0.4 mg plasma proteins with complete
Freund adjuvant at a 1-week interval. The LOU rat chosen for fusion was
boosted by i.v. inoculation of 0.4 mg plasma proteins at days 3,2, and 1
before fusion. Fusion was carried out according to standard procedures in
the Experimental Immunology Unit of the University of Louvain. From the 17
hybridomas selected for their growing properties and their positive
response in an ELISA for vitellogenin, a single one was cloned and the
others frozen for 2 years. After limiting dilution, this hybridoma gave a
monoclonal antibody (MAb) producing hybridoma LO-VG1. This was typed as an
IgG2a immunoglobulin. After 2 years, two other hybridomas were rescued and
provided two MAbs, LO-VG2 and LO-VG3. The three antibodies exhibited
different electrophoretic mobilities. The work with these MAbs was pursued
in two directions: the purification of the MAbs and the setup of an
indirect sandwich ELISA for vitellogenin and antibodies directed against
this antigen.
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