Recombinant Genetic Libraries and Human Monoclonal Antibodies
Recombinant Genetic Libraries and Human Monoclonal Antibodies
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DOI：
10.1007/978-1-62703-586-6_9
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发表时间：
2014-01-01
期刊：
影响因子：
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通讯作者：
Sidhu, Sachdev S.
中科院分区：
文献类型：
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作者：
Adams, Jarrett J.;Nelson, Bryce;Sidhu, Sachdev S.
为了全面操纵人类蛋白质组，我们需要大量的药理学试剂。为了满足这些需求，我们已经通过噬菌体展示开发了合成抗体的库，其中使用多样化的寡核苷酸来修饰人抗原结合片段（Fab）支架的互补决定区（CDR）。由于多样性是在哺乳动物免疫系统的范围之外产生的，合成抗体文库使我们能够绕过杂交瘤技术的几个限制，同时改善生产药理学试剂的实验参数。在这里，我们描述了用于从单一人类框架产生合成抗体文库的方法，其多样性限于四个CDR。这些合成库可以是极其功能性的，因为它们产生对大多数可溶性人抗原的高选择性、高亲和力Fab。最后，我们描述了选择方法，使我们能够克服免疫优势，在我们的选择，以针对每个抗原的各种表位。这些方法使我们能够生产人类单克隆抗体来操纵人类蛋白质组。
In order to comprehensively manipulate the human proteome we require a vast repertoire of pharmacological reagents. To address these needs we have developed repertoires of synthetic antibodies by phage display, where diversified oligonucleotides are used to modify the complementarity-determining regions (CDRs) of a human antigen-binding fragment (Fab) scaffold. As diversity is produced outside the confines of the mammalian immune system, synthetic antibody libraries allow us to bypass several limitations of hybridoma technology while improving the experimental parameters under which pharmacological reagents are produced. Here we describe the methodologies used to produce synthetic antibody libraries from a single human framework with diversity restricted to four CDRs. These synthetic repertoires can be extremely functional as they produce highly selective, high affinity Fabs to the majority of soluble human antigens. Finally we describe selection methodologies that allow us to overcome immuno-dominance in our selections to target a variety of epitopes per antigen. Together these methodologies allow us to produce human monoclonal antibodies to manipulate the human proteome.