本研究通过对鸡败血支原体(MG)强毒株黏附素GapA的N端(98aa-322aa)高变区作为免疫原免疫小鼠.利用Bac-to-bac系统构建GapAN重组杆状病毒,感染Sf9昆虫细胞,以作为检测抗原,筛选获得5株抗GapAN端的单克隆抗体:3D4、5810、185、1D7和4B3。经部分免疫学特性鉴定,5株单抗具备良好的免疫反应性,并且在免疫荧光试验及Western-blot检测过程中发现这些单抗针对的抗原表位存在明显差异,这些单抗的成功制备为下一步深入研究MG的黏附特性奠定了基础。
GapA is known as one of important adhesins in Mycoplasma gallisepticum (MG). The N- terminus of GapA was expressed in prokaryotic system and used as the immune antigen to immunize Balb/C mice. The monoclonal antibodies were screened in ELISA by using the GapA antigen expressed in recombinant Baculovirus infected Sf9 cells. Five monoclonal antibodies specific for GapA were identified and designated as 3D4, 5B10, 1B5, 1D7 and 4B3. Different reaction patterns were observed in indirect florescent (IFA) and Western blot assays, suggesting they might react to diverse epitopes. These antibodies will be useful tools to study the adhesins of Mycoplasma gallisepticum.