Epitope Mapping by Phage Display
Epitope Mapping by Phage Display
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DOI：
10.1007/978-1-4939-7447-4_28
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发表时间：
2018-01-01
期刊：
影响因子：
--
通讯作者：
Hust, Michael
中科院分区：
文献类型：
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作者：
Moreira, Gustavo Marcal Schmidt Garcia;Fuehner, Viola;Hust, Michael
在免疫系统的分子中，抗体，特别是单克隆抗体（mAb），已经显示出对于许多生物学应用是令人感兴趣的。由于它们只能识别其靶标的独特部分，因此mAb通常具有很强的特异性。这些靶标可以有许多不同的组成，但最常见的是通常来自宿主外部的蛋白质或肽，尽管自身蛋白质也可以在自身免疫性疾病或某些类型的癌症中被靶向。与其靶标相互作用的mAb部分组成互补位，而靶标的识别部分组成表位。了解表位对于生物制品的改进是有价值的，例如，诊断测定、治疗性mAb或疫苗，以及用于阐明免疫应答。目前的表位定位技术依赖于靶点或其部分的呈递，其方式是靶点或其部分可以与某种mAb相互作用。尽管有几种技术可供选择，但每种技术都有其优点和缺点。因此，对其中一种技术的选择通常取决于研究人员的偏好和可用性，是否有改进的可能性，或者是否有替代技术的发展。例如，噬菌体展示是一种多功能技术，其允许呈现许多不同的寡肽，这些寡肽可以针对不同的抗体进行测试，从而满足表位作图方法的需要。在本章中，给出了一个用于构建单靶寡肽噬菌体文库的方案，以及用于使用噬菌体展示进行表位作图的淘选程序。
Among the molecules of the immune system, antibodies, particularly monoclonal antibodies (mAbs), have been shown to be interesting for many biological applications. Due to their ability to recognize only a unique part of their target, mAbs are usually very specific. These targets can have many different compositions, but the most common ones are proteins or peptides that are usually from outside the host, although self-proteins can also be targeted in autoimmune diseases, or in some types of cancer. The parts of a mAb that interact with its target compose the paratope, while the recognized parts of the target compose the epitope. Knowing the epitope is valuable for the improvement of a biological product, e.g., a diagnostic assay, a therapeutic mAb, or a vaccine, as well as for the elucidation of immune responses. The current techniques for epitope mapping rely on the presentation of the target, or parts of it, in a way that it can interact with a certain mAb. Even though there are several techniques available, each has its pros and cons. Thus, the choice for one of them is usually dependent on the preference and availability of the researcher, opening possibility for improvement, or development of alternative techniques. Phage display, for example, is a versatile technology, which allows the presentation of many different oligopeptides that can be tested against different antibodies, fitting the need for an epitope mapping approach. In this chapter, a protocol for the construction of a single-target oligopeptide phage library, as well as for the panning procedure for epitope mapping using phage display is given.