相信大家对单克隆抗体 (monoclonal antibodies,mAbs)一定不陌生,而且不少实验需要用到它们。这些抗体通常是通过杂交瘤细胞系统生产出来的。这个方法最早是由 Köhler 和Milstein 在 1975年发明的1。简单来说,就是一只实验小鼠为所需要产生的抗体而进行免疫。将小鼠的脾细胞(spleen cells)-主要由免疫系统中B细胞组成,和特殊的骨髓瘤细胞(myeloma cells)融合,就会产生持续增殖细胞(immortal hybrid cell)。这些细胞被称作杂交瘤细胞(hybridoma),它们可以产生特定的单克隆抗体。
虽然这个技术被广泛使用,但是它同样有许多弊端。为了克服这些问题,重组抗体(recombinant antibodies,rAbs) 应运而生,并且颠覆了整个单克隆抗体的生产模式。那么今儿小编就给大家简单介绍一下rAbs的好处!首先,什么是重组抗体呢?rAbs 是在活生物体之外(in vitro)的实验室中通过基因重组的技术构建的。它们是不受免疫系统的限制的,而且这些重组抗体可以在任何使用mAbs的应用中使用。
那么,重组抗体的好处是什么呢?
1. 它们需要更少的提纯抗原就能被生产出来;
2. 生产的时间从以月份来计量变成以周计量,而且是高通量的(high-throughput);
3. 不像mAbs,rAbs 可以根据任何一种抗原被构建出来,包括一些小鼠免疫系统不能识别的抗原;或是不能激发免疫系统的抗原;和一些对动物体有毒的分子;
4. rAbs 可以防止由小鼠单克隆抗体激发的人类免疫系统反应。mABs,即使是已经“人类化”(humanized)的(图1),也会被人类免疫系统识别,会造成超敏反应(hypersensitivity reaction)。这个反应是免疫反应产生作用分子移除外来抗原的过程。在特殊的状况下,发炎反应可能导致明显的组织伤害甚至死亡。
5. rABs 可以有几种形式:Fab片段,单链变异区片段抗体(single-chain variable region fragments,scFv),双价小抗体(diabodies,dimeric scFvs)等等 (图2)。不但形式多种多样,而且表达的系统也很广泛,例如大肠杆菌,哺乳动物细胞,酵母,昆虫细胞,甚至是植物细胞。
References
1. Köhler G., Milstein C. (1975). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495-97.
2. Patricia A., Herrington-Symes1, Monika Farys1, Hanieh Khalili (2013). Steve Brocchini Antibody fragments: Prolonging circulation half-life special issue-antibody research. Advances in Bioscience and Biotechnology.
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