基因重组人粒细胞集落细胞刺激因子单克隆抗体的制备及特性

Preparation and characteristics of monoclonal antibody against recombinant human granulocyte colony stimulating factor

采用高效免疫方法，利用rhG－CSF免疫BALB／C小鼠，经过两次融合制备了3株抗rhG－CSF的单克隆抗体，分别命名为1H11、2B3、2C3；抗体类型均为IgG2a。免疫印迹试验证明为特异性抗G－CSF的单克隆抗体；单抗相加试验和单抗竞争试验表明3株单抗针对G－CSF两具不同的抗原决定簇；中和试验表明2B、2C3识别的位点与G－CSF刺激NFS－60细胞增殖活性有关。

The research overcame the problems that the G-CSF was a weakly immunogenic antigen to mouse by using the typhoid bacteria body and produced 3 murine hybridoma clones of monoclonal antibodies against recombinant human granuloctye colonystimulating factor by twice cell fusions, The 3 hybridoma clones were named 1H11, 2B3, 2C,. All this antibodies were the IgG2a subclass. Western blot was applied to prove specificity of antibodies against G-CSF. The McAb additivity test and McAb competitive test showed the 2B3 and 2C3 recongized the same epitope of G-CSF and 1H11 recongized another epitope of G-CSF. The neutralization trial of G-CSF McAb revealed 2B3 and 2C3 were capable of neutralizing the proliferative activity of G-CSF to NFS-60 cells.