基于糖和苯硼酸动态共价键的高效蛋白质表面修饰方法

林铭昌; 陈国颂

doi:10.11777/j.issn1000-3304.2017.16347

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基于糖和苯硼酸动态共价键的高效蛋白质表面修饰方法

研究论文 | 更新时间：2021-01-26

- 基于糖和苯硼酸动态共价键的高效蛋白质表面修饰方法
  增强出版
- An Efficient Protein Modification Method Based on Dynamic Covalent Bonding between Sugar and Phenylboronic Acid
- 高分子学报 2017年第7期页码：1113-1120
- 作者机构：
  
  聚合物分子工程国家重点实验室复旦大学高分子科学系上海 200433
- 作者简介：
  
  陈国颂, guosong@fudan.edu.cn Guo-song Chen, E-mail:guosong@fudan.edu.cn
- 基金信息：
  
  国家自然科学基金(基金号91527305);国家自然科学基金(21474020)
- DOI：10.11777/j.issn1000-3304.2017.16347
  中图分类号：
- 纸质出版日期：2017-7，
  
  收稿日期：2016-12-2，
  
  修回日期：2016-12-24，
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林铭昌, 陈国颂. 基于糖和苯硼酸动态共价键的高效蛋白质表面修饰方法[J]. 高分子学报, 2017,(7):1113-1120.

Lin Ming-chang, Chen Guo-song. An Efficient Protein Modification Method Based on Dynamic Covalent Bonding between Sugar and Phenylboronic Acid[J]. Acta Polymerica Sinica, 2017,(7):1113-1120.
林铭昌, 陈国颂. 基于糖和苯硼酸动态共价键的高效蛋白质表面修饰方法[J]. 高分子学报, 2017,(7):1113-1120. DOI： 10.11777/j.issn1000-3304.2017.16347.

Lin Ming-chang, Chen Guo-song. An Efficient Protein Modification Method Based on Dynamic Covalent Bonding between Sugar and Phenylboronic Acid[J]. Acta Polymerica Sinica, 2017,(7):1113-1120. DOI： 10.11777/j.issn1000-3304.2017.16347.

摘要

糖和蛋白质的相互作用参与了很多重要的生命过程.研究糖和蛋白质的相互作用有多种手段，石英晶体微天平（QCM）是其中重要的一种.研究中常需要将蛋白质通过共价键连接在天平芯片的表面.但是，用于检测糖分子的蛋白质多为植物凝集素，它们的分子量大，表面可修饰位点少，通过共价键修饰在芯片表面的效率偏低.本文提出一种基于糖和苯硼酸之间动态共价键的新修饰方法，能够大幅度提高蛋白质在芯片表面的修饰效率.

Abstract

The interaction between sugar and protein occurs in many important biological processes. Several methods can be used to investigate this interaction. Compared to other methods

especially those in solution

quartz crystal microbalance (QCM) can detect interactions on liquid-solid interface

which might be more useful because the proteins are usually mobilized on the chip surface by covalent bond. However

most proteins used in this study belong to plant lectins with high molecular weight and less modification sites. The efficiency of covalent modification of lectins is quite low. In this paper

we propose a new method based on dynamic covalent bond between sugar and phenylboronic acid

which dramatically increases the modification efficiency on surface. On a gold chip of QCM

block copolymer P

BMA-

-PBOB was first coated by spin-coating

glycopolymers (PMan

PGal

PGlc) was then deposited on the surface

via

the dynamic covalent bond between benzoboroxole (BOB) and sugars. The three glycopolymers did not show much differences due to their relatively similar binding ability to BOB-containing polymers. Control experiments proved that the coating of glycopolymers was driven by the dynamic covalent bond. The glycopolymer coating was quite stable even at low pH and high concentration corresponding to monosaccharides. The lectin Concanavalin A (Con A) was immobilized

via

the interaction between Con A and mannopyranoside. It was found that the amount of Con A coated on gold chip

via

the above-mentioned non-covalent method

was much higher than that by the traditional covalent method. By Sauerbrey equation

the amount of Con A deposited

via

non-covalent method was calculated to be 2832 ng/cm

while that of Con A

via

covalent method was only 424.8 ng/cm

. Moreover

on the glycopolymer surface built by non-covalent method

Con A and glyco-micelles could be deposited layer-by-layer continuously

showing the robustness of this non-covalent method. We hope that this method could be further utilized to deposit sugar-coating nanomaterials and even cells with glycocalyx on certain surfaces.

关键词

凝集素分子自组装动态共价键石英晶体微天平

Keywords

LectinMolecular self-assemblyDynamic covalent bondQuartz crystal microbalance

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