摘要
目的制备特异性人肿瘤坏死因子相关凋亡诱导配体单克隆抗体,为建立TRAIL酶联吸附检测方法奠定基础。方法以原核表达系统重组制备的可溶性人sTRAIL作为抗原,利用杂交瘤技术筛选获得抗人TRAIL?mAb杂交瘤细胞株。体内诱生法生产腹水,经间接ELISA方法检测腹水效价,Western blot鉴定抗体特异性。结果制备获得一株可稳定分泌抗人TRAIL mAb的杂交瘤细胞株,属IgG2b亚类;腹水效价达1:5×10^6以上,Western blot鉴定结果显示可特异性识别人TRAIL,而与TNF-α和Fas-l无明显的交叉反应。结论制备了具有高特异抗原结合特性的TRAIL mAb。
Objective:To prepare and characterize the monoclonal antibody against human TRAIL. Methods:Recombinant soluble sTRAIL was produced in Escherichia coli and purified by affinity chromatography and then subsequently used as immunogen to screen antibodies produced by hybridoma cells. The titers of mAbs were measured by indirect ELISA and the specificities of the m Abs was evaluated by western blot assays. Results:One hybridoma cell line which secrete mAbs against TRAIL were screened and their immunoglobulin subclasses were IgG2 b. The ELISA titer of the ascites fluid was 1:5×10^6 and westerm blot analysis confirmed that the mAb could reacted with recombinant TRAIL with good sensitivity. Conclusion:The mAb against human TRAIL were successfully prepared.
作者
周岩
武坚锐
孙夏瑜
ZHOU Yan;WU Jian-lai;SUN Xia-yu(Shanxi Children's Hospital,Shanxi Maternal and Child Health Care Hospital,Shanxi Taiyuan 030013)
出处
《中国优生与遗传杂志》
2018年第12期14-15,25,共3页
Chinese Journal of Birth Health & Heredity
