抗人IgG重链单克隆抗体和抗人IgG亚类限制性单克隆抗体的鉴定

本文对前获得的4株针对人IgG重链杂交瘤细胞株(1B3,2D5,2A4和2B6)所分泌的单克隆抗体(McAbs)用琼脂糖免疫双扩散试验进行初步分析,发现该4种McAbs是针对人IgG上的三个不同抗原决定簇.利用ELISA竞争试验得到进一步证实.用人IgG亚类免疫球蛋白分析4种McAbs,证明1B3 McAb可与IgG的所有4个亚类反应.其他3种 McAbs为 IgG亚类限制性 McAbs,2B6和 2D5不与 IgG4反应,称缺-IgG4 McAb; 2A4不与 IgG3反应,称缺-IgG3 McAb. 在琼脂糖免疫双扩散试验证实:(1)单独一种McAb 与抗原反应仅能出现透明沉淀线,但在琼脂糖中加入2% PEG 6000可出现常规所见的乳白色沉淀线;(2)将两个McAbs加入同一抗体孔内与抗原反应,当两个McAbs不同时(如2A4与2D5)则出现乳白色沉淀线.上述结果提示可用乳白色沉淀线的出现与否来初步鉴别两种McAbs的异同. 以上亚类限制性McAbs,在未获得亚类特异性McAb以前,可用于制备和纯化人 IgG3和 IgG4,亦可用于研究人 IgG的抗原分析。     

IDENTIFICATION OF ANTI-HUMAN IgG MONOCLONAL ANTIBODIES AND MONOCLONAL ANTIBODIES WITH CONFINED SUBCLASS REACTIVITY

In a previous paper, 4 hybridoma cell lines (1B3, 2D5, 2A4 and 2B6) secreting antibodiesagainst heavy chain of hmnan IgG were reported. In this paper a more detailed study of theseMcAbs was introduced. First of all they were studied with double immunodiffusion test inagarose and it was demonstrated that they were reactive against 3 distinct antigenic deter-minants of human IgG. This result was also confirmed by ELISA competition test. TheseMeAbs were further analysed for human IgG subclass proteins and it was found that 1B3McAb gave positive reaction with all subclasses of IgG while the other 3 possessed confinedsubclass reactivity, i.e. McAb 2A4 did not react with IgG3 (ealled non-IgG3 McAb), andboth McAbs 2D5 and 2B6 did not react with IgG4 (called non-IgG4 McAb). By agarose immunodiffusion test it was shown that: (i) Only transparent precipitation linewas formed when McAb reacted with IgG antigen, however, in agarose containing 2% PEG-6000, the transparent precipitation line becomes opaque; (ii) When