Lee Jihwan, Lee Suhyun, Park Younbae, Lee Seokhyun, Ha Seungmin, Han Manhye, Jang Gulwon, Park Myunghum, Kim Kyungwoon, Chung Hakjae
Dairy Science Division, National Institute of Animal Science, RDA, Cheonan-si 31000, Korea.
Animal Breeding & Genetics Division, National Institute of Animal Science, RDA, Cheonan-si 31000, Korea.
Animals (Basel). 2021 May 28;11(6):1582. doi: 10.3390/ani11061582.
To improve reproductive performance in cattle, the accurate detection of estrus and optimization of insemination relative to ovulation are necessary. However, poor heat detection by farm staff leads to a decreased conception rate, thus inflicting economic damage to the beef and dairy industries. This study aimed to develop monoclonal antibodies (mAb) that can specifically bind to the bovine lactoferrin (bLF) protein, which we have previously demonstrated to be overexpressed in bovine cervical mucus during estrus. Female rats were intraperitoneally immunized with bLF protein as the antigen. Anti-bLF mAbs were then purified by affinity chromatography, and their binding affinity for the bLF antigen was examined using ELISA. We found a high binding affinity between mAbs and bLF. Finally, we developed a rapid bovine heat detection kit using the anti-bLF mAbs that we generated and tested on cervical mucus from 12 cows (estrous synchronization, = 2; natural cycling, = 10). We found that the kits accurately detected estrus. Overall, our fabricated heat detection kit based on rat anti-bLF mAbs could pave the way for the development of potent tools for heat detection devices for dairy cattle, thereby preventing economic loss.
为提高牛的繁殖性能，准确检测发情期并优化授精与排卵的相关性十分必要。然而，农场工作人员对发情的检测不佳会导致受孕率下降，从而给牛肉和乳制品行业造成经济损失。本研究旨在开发能够特异性结合牛乳铁蛋白（bLF）的单克隆抗体（mAb），我们之前已证明该蛋白在发情期的牛宫颈黏液中过表达。以bLF蛋白为抗原对雌性大鼠进行腹腔免疫。然后通过亲和层析纯化抗bLF单克隆抗体，并使用酶联免疫吸附测定（ELISA）检测它们与bLF抗原的结合亲和力。我们发现单克隆抗体与bLF之间具有高结合亲和力。最后，我们使用所产生的抗bLF单克隆抗体制备了一种快速牛发情检测试剂盒，并在12头奶牛（发情同步，n = 2；自然发情周期，n = 10）的宫颈黏液上进行了测试。我们发现该试剂盒能准确检测出发情期。总体而言，我们基于大鼠抗bLF单克隆抗体制备的发情检测试剂盒可为开发高效的奶牛发情检测设备工具铺平道路，从而避免经济损失。
