褪黑素（MT）广泛存在于哺乳动物体内,是一种常见的吲哚类物质,可起到清除自由基、调节免疫、调节神经等作用。本文使用褪黑素-OVA作为免疫原,免疫5 只BALB/c小鼠,5 次免疫后采血检测,通过间接ELISA方法确定抗血清针对褪黑素-OVA的滴度。经ELISA筛选,最终获得5 株可稳定表达褪黑素抗体的单克隆细胞株（3G3B2、3G3C7、3G3D1、3G3F4、3G3G3）。纯化后抗体浓度为0.28 mg/mL,经测定,褪黑素单克隆抗体滴度可达1∶64 000,为进一步开发褪黑素的快速检测方法奠定了材料基础。
Melatonin （MT）,which is widely existed in mammals,is a common indole substance that can play a role in scavenging free radicals,regulating immunity,and regulating nerves. Here,five BALB/c mice were immunized with melatonin -OVA as the immunogen,and blood was collected for detection after five immunizations. The titer of antiserum against melatonin-OVA was determined by indirect ELISA. After ELISA screening,5 monoclonal cell lines（3G3B2,3G3C7,3G3D1,3G3F4 and 3G3G3） stably expressing melatonin antibody were obtained. The concentration of purified antibody was 0.28 mg/mL,and the titer of purified antibody reached 1∶64 000. The melatonin monoclonal antibody produced in this research laid a material foundation for further development of a rapid detection method of melatonin.
[1] Carrillo-Vico A,Lardone PJ,Álvarez-Sánchez N,et al.Melatonin:buffering the immune system[J].International Journal of Molecular Sciences,2013,14(4):8638-8683.
[2] Tamura H,Kawamoto M,Sato S,et al.Long-term melatonin treatment delays ovarian aging[J].Journal of Pineal Research,2017,62:e12381.
[3] Mortezaee K,Potes Y,Mirtavoos-Mahyari H,et al.Boosting immune system against cancer by melatonin:a mechanistic viewpoint[J].Life Sciences,2019,238:116960-116960.
[4] Kim H J,Kim H J,Bae M K,et al.Suppression of osteoclastogenesisby melatonin:amelatonin receptor-independent action[J].International Journal of Molecular Sciences,2017,18(6):1142.
[5] Jones LH.Recent advances in the molecular design of synthetic vaccines[J]. Nature Chemistry,2015,7(12):952-60.
[6] 杨松鑫,冯建远,郭旋,等.小分子化合物单克隆抗体的制备及其在动物医学领域的应用[J].中国畜牧兽医,2021,48(3):1121-1131.
[7] Wang H,Wang H,Huang H,et al.Melatonin attenuates spinal cord injury in mice by activating the nrf2/are signaling pathway to inhibit the nlrp3 inflammasome[J]. Cells,2022,11(18):2809.
[8] Jiang Y H,Shi H C,Liu Y,et al,Applications of melatonin in female reproduction in the context of oxidative stress[J],Oxidative Medicine and Cellular Longevity,2021,e6668365.
