Nanoyeast and Other Cell Envelope Compositions for Protein Studies and Biosensor Applications.
Nanoyeast and Other Cell Envelope Compositions for Protein Studies and Biosensor Applications.
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DOI：
10.1021/acsami.6b09263
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发表时间：
2016-11-16
影响因子：
9.5
通讯作者：
Trau, Matt
中科院分区：
文献类型：
--
作者：
Grewal, Yadveer S.;Shiddiky, Muhammad J. A.;Mahler, Stephen M.;Cangelosi, Gerard A.;Trau, Matt
疾病生物标志物发现的快速进展增加了对强大检测技术的需求。在过去的几年中，许多免疫亲和试剂的设计重点是降低成本和提高特异性，同时提高稳定性。抗体片段（scFv）早已展示在酵母和噬菌体文库的表面以供选择；然而，此类片段的稳定生产带来了挑战，阻碍了它们在诊断中的广泛使用。当从天然环境中溶解时，膜和细胞壁蛋白同样会遇到稳定性问题。最近，已经开发了将膜蛋白维持在天然或类天然脂质环境中以提高其稳定性的细胞包膜组合物。这种细胞包膜组成方法现已适应通过保留其天然细胞壁环境来稳定抗体片段。开发出一类新型免疫亲和试剂，可维持抗体片段与酵母细胞壁的附着。在此，我们回顾了将细胞壁片段与功能性 scFv 相结合的最新策略，这些策略旨在轻松生产，同时在与简单的检测平台一起使用时保持特异性和稳定性。这些基于细胞壁的抗体片段结构呈球状，大小不一，片段大小从几十到几百纳米不等。一旦固定到生物传感器表面上，这些片段似乎保留了活性，用于特异性和灵敏地检测病原体抗原。它们可以快速、经济地从酵母展示库中产生，并在室温下冻干保存长达一年，对稳定性影响很小。这种新形式的 scFv 以简单且低成本的方式为 scFv 在生物传感器应用中的使用提供了稳定性。这种抗体细胞壁复合材料的生产和“淘选”也非常容易，使得能够快速采用稳定且廉价的亲和试剂来应对新出现的感染威胁。
Rapid progress in disease biomarker discovery has increased the need for robust detection technologies. In the past several years, the designs of many immunoaffinity reagents have focused on lowering costs and improving specificity while also promoting stability. Antibody fragments (scFvs) have long been displayed on the surface of yeast and phage libraries for selection; however, the stable production of such fragments presents challenges that hamper their widespread use in diagnostics. Membrane and cell wall proteins similarly suffer from stability problems when solubilized from their native environment. Recently, cell envelope compositions that maintain membrane proteins in native or native-like lipid environment to improve their stability have been developed. This cell envelope composition approach has now been adapted toward stabilizing antibody fragments by retaining their native cell wall environment. A new class of immunoaffinity reagents has been developed that maintains antibody fragment attachment to yeast cell wall. Herein, we review recent strategies that incorporate cell wall fragments with functional scFvs, which are designed for easy production while maintaining specificity and stability when in use with simple detection platforms. These cell wall based antibody fragments are globular in structure, and heterogeneous in size, with fragments ranging from tens to hundreds of nanometers in size. These fragments appear to retain activity once immobilized onto biosensor surfaces for the specific and sensitive detection of pathogen antigens. They can be quickly and economically generated from a yeast display library and stored lyophilized, at room temperature, for up to a year with little effect on stability. This new format of scFvs provides stability, in a simple and low-cost manner toward the use of scFvs in biosensor applications. The production and “panning” of such antibody cell wall composites are also extremely facile, enabling the rapid adoption of stable and inexpensive affinity reagents for emerging infectious threats.
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影响因子：
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作者：
Ahmad ZA;Yeap SK;Ali AM;Ho WY;Alitheen NB;Hamid M
通讯作者：
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影响因子：
5.6
作者：
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影响因子：
2.4
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通讯作者：
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作者：
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通讯作者：
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