MacDonald G, Primrose S, Biggins K, Bowman J M, Berczi I, Friesen A D, Sehon A H
Scand J Immunol. 1987 May;25(5):477-83. doi: 10.1111/j.1365-3083.1987.tb02219.x.
Human monoclonal antibodies specific for the Rh(D) antigen were produced by cell lines generated by the fusion of pooled Epstein-Barr virus (EBV)-transformed B-cell lines secreting Rh(D) antibodies with the murine myeloma cell line NS.1 or with the human lymphoblastoid cell line HOA.1. The selection of hybrids was achieved in RPMI 1640 medium containing HAT and ouabain. Higher fusion efficiency was obtained with the NS.1 cell line; however, the hybrids with HOA.1 exhibited a greater clonal stability. The products of four clones (three human-human and one human-mouse) that consistently secreted antibodies for over 11 months were tested for specificity with a panel of red cells of various Rh phenotypes. The supernatants of all four clones showed anti-Rh(D) specificity but failed to react with the red cell Du phenotypes categorized as DV(Dw+) and DVI. Two of the three human-human clones secreted IgM(lambda) and the third IgG(kappa). The human-mouse clone produced IgG(kappa) antibody.
针对Rh(D)抗原的人源单克隆抗体是由汇集的分泌Rh(D)抗体的爱泼斯坦-巴尔病毒(EBV)转化B细胞系与鼠骨髓瘤细胞系NS.1或人淋巴母细胞系HOA.1融合产生的细胞系所产生。杂种细胞的选择是在含有次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷(HAT)和哇巴因的RPMI 1640培养基中进行的。用NS.1细胞系获得了更高的融合效率；然而，与HOA.1的杂种细胞表现出更高的克隆稳定性。对持续分泌抗体超过11个月的四个克隆(三个是人-人杂交克隆和一个是人-鼠杂交克隆)的产物，用一组具有不同Rh表型的红细胞检测其特异性。所有四个克隆的上清液均显示抗Rh(D)特异性，但不与分类为DV(Dw+)和DVI的Du红细胞表型发生反应。三个是人-人杂交克隆中的两个分泌IgM(λ)，第三个分泌IgG(κ)。人-鼠杂交克隆产生IgG(κ)抗体。
