类弹性蛋白作为功能纳米材料在生物工程领域研究及应用进展

doi:10.13560/j.cnki.biotech.bull.1985.2020-0042

摘要/Abstract

摘要：

类弹性蛋白（Elastin-like polypeptide,ELP）是一种具有弹性功能且对温度敏感的人造蛋白质聚合物,主要由五肽重复序列单元Val-Pro-Gly-Xaa-Gly（VPGXG）_n构成。ELP对温度敏感的可逆相变特征体现在低于相变温度（Transition temperature,Tt）时呈可溶状态,在高于相变温度的溶液中呈凝聚状态,此过程可逆。基于ELP标签的可逆相变循环（Inverse transition cycle,ITC）是一种新型的分离纯化重组蛋白质的非色谱技术。利用ELP融合蛋白的可逆相变特性,经过多次ITC循环从可溶的蛋白上清溶液中特异性分离出含有ELP标签的融合蛋白。该技术仅需常规的离心机就能得到与色谱法的亲和层析纯度相当的目的蛋白,成本低、操作简便,适合大规模生产的需要。ELP和ELP衍生分子具有良好的弹性、自组装性、长期稳定性和生物活性等特性,精确控制ELP的设计有利于操纵它们对外界环境的刺激反应和其他物理功能特征,ELP可调的理化性质和良好生物相容性使ELP在生物学上也具有广泛的应用。重点阐述了ELP相变特性及机理,ELP融合蛋白的设计,ELP标签对融合蛋白的影响,ELP标签在蛋白质纯化、生物医药、生物工程、水凝胶及新兴领域方面的应用,ELP发展方向及亟待解决的关键问题和技术。

关键词: 类弹性蛋白, ITC技术, ELP标签, 蛋白纯化, 生物应用

Abstract:

Elastin-like polypeptides（ELP）are an elastic and temperature-sensitive synthetic protein polymer composed of a Val-Pro-Gly-Xaa-Gly repeat unit derived from human tropoelastin. ELPs undergo a sharp reversible phase transition at specific temperatures known as the inverse transition temperature（T_t）. Below its T_t,ELP assumes a random coil shape,is highly solvated and hence highly soluble in aqueous solution. However,when temperature is raised above its T_t,the ELP becomes insoluble,forms large microns and begins to aggregate. ELP-based inverse transition cycle（ITC）is a non-chromatographic technique for the separation and purification of recombinant protein. Using the reversible phase change characteristics of ELP fusion proteins,ELP-labeled fusion proteins are specifically isolated from soluble protein supernatant after multiple ITC cycles. This technique requires only conventional centrifuges to obtain the target protein of the same purity as that by affinity chromatography,and it is low cost,easy to operate and suitable for lage-scale production. ELP and ELP-derived molecules present properties of fine elasticity,self-assembly,long-term stability and biological activity. The precise control in designing ELP is conducive to manipulating their stimulation responses to the external environment and other physical functional characteristics. The adjustable physical and chemical properties of ELP and its promising biocompatibility make ELP widely used in biotechnology. In this paper,the phase transition characteristics and mechanism of ELP,the design of ELP fusion protein and the effect of ELP to fusion proteins（peptides）are reviewed. Meanwhile,the application of ELP tag in protein purification,biomedicine and bioengineering,hydrogels and emerging fields is reviewed. Finally,the research directions and key issues and technologies of ELP also are discussed.

Key words: elastin-like polypeptide, ITC technology, ELP tag, protein purification, biological application

周阳, 王桃桃, 闫丹丹, 王莹莹, 施沁璇, 孙丽慧, 林锋. 类弹性蛋白作为功能纳米材料在生物工程领域研究及应用进展[J]. 生物技术通报, 2020, 36(11): 198-208.

ZHOU Yang, WANG Tao-tao, YAN Dan-dan, WANG Ying-ying, SHI Qin-xuan, SUN Li-hui, LIN Feng. Advances in Biotechnological Application of Elastin-like Polypeptides as Functional Nanomaterials[J]. Biotechnology Bulletin, 2020, 36(11): 198-208.

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