RABBIT MONOCLONAL-ANTIBODIES - GENERATING A FUSION PARTNER TO PRODUCE RABBIT RABBIT HYBRIDOMAS
RABBIT MONOCLONAL-ANTIBODIES - GENERATING A FUSION PARTNER TO PRODUCE RABBIT RABBIT HYBRIDOMAS
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DOI：
10.1073/pnas.92.20.9348
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发表时间：
1995-09-26
影响因子：
11.1
通讯作者：
KNIGHT, KL
中科院分区：
文献类型：
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作者：
SPIEKERPOLET, H;SETHUPATHI, P;KNIGHT, KL
在过去 15 年中，多个实验室尝试生产兔单克隆抗体，主要是因为兔识别小鼠或大鼠中不具有免疫原性的抗原和表位，而这两种物种通常是产生单克隆抗体的。然而，由于无法获得浆细胞瘤融合配偶体，因此无法产生来自兔子的单克隆抗体。为了获得可用作融合伴侣的兔浆细胞瘤细胞系，我们产生了携带两种转基因c-myc和v-abl的转基因兔。这些兔子出现了浆细胞瘤，我们获得了几种浆细胞瘤细胞系，从中分离出次黄嘌呤/氨基蝶呤/胸苷敏感克隆。其中一个克隆与免疫兔子的脾细胞融合时，产生稳定的杂交瘤，分泌针对免疫原的特异性抗体。杂交瘤可以在裸鼠中克隆和增殖，并且可以将它们冷冻而不改变其分泌特异性单克隆抗体的能力。这些兔-兔杂交瘤不仅可用于单克隆抗体的生产，还可用于免疫球蛋白基因重排和同种型转换的研究。
During the last 15 years several laboratories have attempted to generate rabbit monoclonal antibodies, mainly because rabbits recognize antigens and epitopes that are not immunogenic in mice or rats, two species from which monoclonal antibodies are usually generated. Monoclonal antibodies from rabbits could not be generated, however, because a plasmacytoma fusion partner was not available. To obtain a rabbit plasmacytoma cell line that could be used as a fusion partner we generated transgenic rabbits carrying two transgenes, c-myc and v-abl. These rabbits developed plasmacytomas, and we obtained several plasmacytoma cell lines from which we isolated hypoxanthine/aminopterin/thymidine-sensitive clones. One of these clones, when fused with spleen cells of immunized rabbits, produced stable hybridomas that secreted antibodies specific for the immunogen. The hybridomas can be cloned and propagated in nude mice, and they can be frozen without change in their ability to secrete specific monoclonal antibodies. These rabbit-rabbit hybridomas will be useful not only for production of monoclonal antibodies but also for studies of immunoglobulin gene rearrangements and isotype switching.