Use of monoclonal antibodies for radioimmunoassay of water buffalo milk progesterone.
In order to standardize a radioimmunoassay of milk progesterone as a routine method for confirmation of oestrus and diagnosis of pregnancy in water buffalo, monoclonal antibodies against progesterone were produced. Hybridomas were prepared by fusing spleen cells from a Balb/c mouse immunized with progesterone 11 alpha-hemisuccinate-bovine serum albumin conjugate with the mouse myeloma cell line NS-1. Thirty wells out of 94 secreted anti-progesterone antibodies. Of the ten independent hybridomas derived, one (AF65) was suitable for the quantification of milk progesterone by radioimmunoassay. The tracer used in the assay was progesterone-11 alpha-hemisuccinate [( 2-125I]iodohistamine). The sensitivity of the assay was 50 pg/tube. The mean progesterone concentration at oestrus was 0.8 +/- 0.2 ng/ml increasing to 8.5 +/- 0.8 ng/ml 24 d later in pregnant animals.
Institute of Animal Production, University of Naples, Italy.
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R
Capparelli
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Iannelli
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Bordi
D000818:Animals; D000911:Antibodies, Monoclonal; D001947:Breeding / Q000379:methods*; D002020:Buffaloes / Q000502:physiology*; D004972:Estrus Detection / Q000379:methods*; D005260:Female; D008892:Milk / Q000032:analysis*; D011247:Pregnancy; D011258:Pregnancy Tests / Q000379:methods / Q000662:veterinary*; D011374:Progesterone / Q000032:analysis*; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D011374:Progesterone