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评估含辅因子生物活性酶的上转换磷光体的共价连接和能量转移能力。

Assessing the Covalent Attachment and Energy Transfer Capabilities of Upconverting Phosphors With Cofactor Containing Bioactive Enzymes.

作者信息

Burgess Letitia, Wilson Hannah, Jones Alex R, Hay Sam, Natrajan Louise S

机构信息

Department of Chemistry, School of Natural Sciences, The University of Manchester, Manchester, United Kingdom.

Manchester Institute of Biotechnology, The University of Manchester, Manchester, United Kingdom.

出版信息

Front Chem. 2020 Dec 21;8:613334. doi: 10.3389/fchem.2020.613334. eCollection 2020.

DOI:10.3389/fchem.2020.613334
PMID:33409268
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7779683/
Abstract

Upconverting phosphors (UCPs) convert multiple low energy photons into higher energy emission via the process of photon upconversion and offer an attractive alternative to organic fluorophores for use as luminescent probes. Examples of biosensors utilizing the apparent energy transfer of UCPs and nanophosphors (UCNPs) with biomolecules have started to appear in the literature but very few exploit the covalent anchoring of the biomolecule to the surface of the UCP to improve the sensitivity of the systems. Here, we demonstrate a robust and versatile method for the covalent attachment of biomolecules to the surface of a variety of UCPs and UCNPs in which the UCPs were capped with functionalized silica in order to provide a surface to covalently conjugate biomolecules with surface-accessible cysteines. Variants of BM3Heme, cytochrome C, glucose oxidase, and glutathione reductase were then attached via maleimide-thiol coupling. BM3Heme, glucose oxidase, and glutathione reductase were shown to retain their activity when coupled to the UCPs potentially opening up opportunities for biosensing applications.

摘要

上转换荧光粉（UCPs）通过光子上转换过程将多个低能光子转换为更高能量的发射，为用作发光探针的有机荧光团提供了一种有吸引力的替代方案。利用UCPs和纳米荧光粉（UCNPs）与生物分子之间明显的能量转移的生物传感器实例已开始出现在文献中，但很少有利用生物分子与UCP表面的共价锚定来提高系统灵敏度的。在此，我们展示了一种强大且通用的方法，用于将生物分子共价连接到各种UCPs和UCNPs的表面，其中UCPs用功能化二氧化硅包覆，以便提供一个表面，使生物分子能够与表面可及的半胱氨酸共价共轭。然后通过马来酰亚胺 - 硫醇偶联连接BM3血红素、细胞色素C、葡萄糖氧化酶和谷胱甘肽还原酶的变体。当与UCPs偶联时，BM3血红素、葡萄糖氧化酶和谷胱甘肽还原酶显示出保留其活性，这可能为生物传感应用开辟机会。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/3116559d39ca/fchem-08-613334-g0011.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/fa3490c5207e/fchem-08-613334-g0013.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/8c7b11322620/fchem-08-613334-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/b6ebefb57841/fchem-08-613334-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/eefc224b5f57/fchem-08-613334-g0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/bb287b26c47c/fchem-08-613334-g0008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/59744825b173/fchem-08-613334-g0009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/0bc095eb07c3/fchem-08-613334-g0010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/3116559d39ca/fchem-08-613334-g0011.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/63ab36a003af/fchem-08-613334-g0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/ff00af3df3c6/fchem-08-613334-g0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/732bc9234bfc/fchem-08-613334-g0012.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/ffb0bf7d5b43/fchem-08-613334-g0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/f9f94ea0e880/fchem-08-613334-g0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/fa3490c5207e/fchem-08-613334-g0013.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/8c7b11322620/fchem-08-613334-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/b6ebefb57841/fchem-08-613334-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/eefc224b5f57/fchem-08-613334-g0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/bb287b26c47c/fchem-08-613334-g0008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/59744825b173/fchem-08-613334-g0009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/0bc095eb07c3/fchem-08-613334-g0010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8a7/7779683/3116559d39ca/fchem-08-613334-g0011.jpg

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