来自护士鲨的单结构域V区文库的生成与筛选。
The generation and selection of single-domain, v region libraries from nurse sharks.
作者信息
Flajnik Martin F, Dooley Helen
机构信息
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
出版信息
Methods Mol Biol. 2009;562:71-82. doi: 10.1007/978-1-60327-302-2_6.
The cartilaginous fish (sharks, skates, and rays) are the oldest phylogenetic group in which a human-type adaptive immune system and immunoglobulins (Igs) have been found. In addition to their conventional (heavy-light chain heterodimeric) isotypes, IgM and IgW, sharks produce the novel isotype, IgNAR, a heavy chain homodimer that does not associate with light chains. Instead, its variable (V) regions act as independent, soluble units in order to bind antigen. In this chapter, we detail our immunization protocol in order to raise a humoral IgNAR response in the nurse shark (Ginglymostoma cirratum) and the subsequent cloning of the single-domain V regions from this isotype in order to select antigen-specific binders by phage display.
软骨鱼（鲨鱼、鳐鱼和魟鱼）是发现人类类型适应性免疫系统和免疫球蛋白（Ig）的最古老的系统发育群体。除了其传统的（重链-轻链异源二聚体）同种型IgM和IgW外，鲨鱼还产生新型同种型IgNAR，这是一种不与轻链结合的重链同型二聚体。相反，其可变（V）区作为独立的可溶性单位来结合抗原。在本章中，我们详细介绍了免疫方案，以在护士鲨（Ginglymostoma cirratum）中引发体液IgNAR反应，以及随后从该同种型中克隆单域V区，以便通过噬菌体展示选择抗原特异性结合物。
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