Head (H) And Tail (T) Monoclonal Antibodies of Boar Spermatozoa

Abstract

Semen collected from three boars were pooled and the sperm cells separated from the semen, washed, sonicated and mixed with equal volume of Freund's adjuvant. The mixture of sperm with complete Freund's adjuvant was used to immunize five mice of the BALB/C strain intraperitoneally on Day O. The sonicated sperm cell immunize the mice the same tourte on Day 14. The mice were immunized with suspension in Incomplete Freund's adjuvant was used to sonicated sperm without adjuvant on Day 21 before they were sacrificed on Day 24. Spleen cells prepared from the mice were fused with myeloma cells (Ag.8.653) by addition of polyethylene glycol (PEG). Two distinct monoclonal antibodies (antihead (H) and anti-tail (T) Mabs) were produced after cloning of the hybridoma cells. There were intermediate forms that are believed to have arisen as a result of incomplete cloning. The Mabs were identified by the use of indirect immunofluorescence method and their specificity confirmed by their non-reaction with cells from some other organs. These Mabs can be used for further work on the mechanisms of sperm antibody effect on swine infertility. It is speculated that the H. Mab and the T Mab may block acrosome reaction and sperm motility respectively. Keyword: Boar spermatozoa monoclonal antibodies Trop. Vet. Vol. 21 (2) 27-34 (2003)