Capparelli R, Iannelli D, Bordi A
Institute of Animal Production, University of Naples, Italy.
J Dairy Res. 1987 Nov;54(4):471-7. doi: 10.1017/s002202990002567x.
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.
为了规范牛奶孕酮放射免疫分析方法，使其成为确认水牛发情和诊断妊娠的常规方法，制备了抗孕酮单克隆抗体。通过将用孕酮11α-半琥珀酸-牛血清白蛋白偶联物免疫的Balb/c小鼠的脾细胞与小鼠骨髓瘤细胞系NS-1融合，制备杂交瘤。94个孔中有30个分泌抗孕酮抗体。在衍生的10个独立杂交瘤中，有一个（AF65）适用于通过放射免疫分析定量牛奶中的孕酮。该分析中使用的示踪剂是孕酮-11α-半琥珀酸[（2-125I]碘组胺）。该分析的灵敏度为50 pg/管。发情期孕酮的平均浓度为0.8±0.2 ng/ml，怀孕动物在24天后增加到8.5±0.8 ng/ml。
