摘要
目的 :获得有生物活性的小鼠抗重组人白细胞介素 18(rhIL 18)单克隆抗体。方法 :采用重组hIL 18免疫BALB C小鼠 ,应用杂交瘤技术 ,ELISA法筛选阳性杂交瘤细胞株并经多次克隆化。结果 :建立了 2株小鼠抗hIL 18单克隆抗体杂交瘤细胞 1C7和 1F5 ,染色体数目分别为 92条和 90条 ,所分泌的抗体分别为IgG2a和IgG1,轻链均为κ型 ,腹水IgG抗体经亲和层析法纯化后纯度达 95 %以上 ,效价为 1× 10 - 4和 1× 10 - 5,Westernblot显示 2株单抗均能特异性识别 18 3kD处的rhIL 18蛋白。 1C7单抗的亲和常数Ka=1 7× 10 5,1F5的亲和常数Ka=1 3× 10 5。 2株单抗识别不同抗原表位。结论 :制备的 2株单抗为进一步研究IL 18的分子结构、生物学功能及其与免疫相关性疾病的关系提供了实验材料。
Objective:To produce hybridoma cell lines of monoclonal antibodies against recombinant human interleukin 18.Methods:Hybridoma techniques were used to fuse BALB/C mice myeloma cell line SP2/0 with B lymphocytes of spleen of BALB/C mice which were immunostimulated with interleukin 18 antigen through polyethyleneglycol (PEG).Single cell lines which stably produce antibodies against interleukin 18 were picked out and identified by enzyme linked immunosorbent assay.Results:Two hybridoma cell lines were obtained and named 1C7 and 1F5,which produce monoclonal antibodies specificity against interleukin 18.They respectively own 88 and 92 chromosomes, the purities of IgG in ascites are all over 95% through affinity chromatography, the titer degrees of ascies are 1×10 -4 and 1×10 -5 respectively.Two antibodies are against two different antigen binding epitopes on the surface of interleukin 18.It was evaluated by Western blot that the two antibodies can bind to interleukin 18 specifically and are made of only single protein strap.Conclusion:Have prepared two hybridoma cell lines which stably secrete high titer and high specific monoclonal antibodies against interleukin 18,which provide the foundation for further studying the structure and the biological function of interleukin 18.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2003年第6期412-414,417,共4页
Chinese Journal of Immunology
