Preparation of Boronic Acid-Functionalized Mesoporous Nanomaterial and Its Application in Enrichment of Glycopeptides
Preparation of Boronic Acid-Functionalized Mesoporous Nanomaterial and Its Application in Enrichment of Glycopeptides
复制标题
硼酸功能化介孔纳米材料的制备及其在糖肽富集中的应用
DOI：
10.6023/a13020176
复制
发表时间：
2013
影响因子：
2.5
通讯作者：
Lu Haojie
中科院分区：
文献类型：
--
作者：
Liu Liting;Zhang Ying;Jiao Jing;Lu Haojie
采用两步后接枝法制备了硼酸功能化介孔纳米材料。首先，将3-缩水甘油氧基丙基三甲氧基硅烷（GLYMO）与3-氨基苯基硼酸一水合物（APB）在氢氧钠溶液（pH 9.18）中反应，制备硼酸键合的GLYMO（记为GLYMO-APB）。其次，将MCM-41添加到制备好的GLYMO-APB溶液中，制备硼酸功能化MCM-41的最终产物（记为MCM-41-GLYMO-APB）。通过 FT-IR 对所制备的材料进行了表征，结果表明硼酸基团成功接枝到 MCM-41 上。基于聚糖链上硼羟基与顺式二醇之间形成环状二酯，可以高选择性和高效率地富集糖肽。对MCM-41-GLYMO-APB富集标准蛋白混合酶切物中糖肽的条件进行了比较和优化。对富集糖肽的孵育时间、孵育溶液、洗涤方式和洗脱液进行了研究。当使用碳酸氢铵缓冲液（100 mmol/L，pH 8.0）作为孵育溶液并孵育1 h时，可以获得最大数量的辣根过氧化物酶（HRP）消化物中的富集糖肽。富集后，用孵育液洗涤制备的材料两次，每次5分钟，可以很大程度上去除非糖肽干扰。当使用含有2,5-二羟基苯甲酸的1%三氟乙酸作为洗脱液时，结合在MCM-41-GLYMO-APB表面的富集糖肽可以被释放，使用这种优化的洗脱液时可以简化操作步骤，因为之后不需要添加MALDI基质溶液。因此，该优化方案为硼酸功能化材料富集糖肽提供了参考，也为利用硼酸化学方法富集生物样品中糖肽的研究奠定了基础。同时，MCM-41-GLYMO-APB高度有序的六角柱状介孔结构为用于生物样品中糖肽组的研究提供了可能。关键词 糖蛋白；硼酸功能化介孔纳米材料；丰富;质谱分析
A boronic acid-functionalized mesoporous nanomaterial was prepared by using a two-step post-grafting method. Firstly, the 3-glycidyloxypropyltrimethoxysilane (GLYMO) and 3-aminophenylboronic acid monohydrate (APB) were re- acted in oxyhydrogen sodium solution (pH 9.18) to prepare boronic-acid bonded GLYMO (denoted as GLYMO-APB). Sec- ondly, the MCM-41 was added into the prepared GLYMO-APB solution to prepare the final product of boronic acid func- tionalized MCM-41 (denoted as MCM-41-GLYMO-APB). This as-prepared material was characterized by FT-IR and the results showed that the boronic acid groups were successfully grafted to MCM-41. Glycopeptides can be enriched with high selectivity and high efficiency based on the formation of a cyclic diester between boronic hydroxyl and cis-diol on the glycan chain. The conditions of enriching glycopeptides in mixed digests of standard proteins by MCM-41-GLYMO-APB were compared and optimized. The incubation time, incubation solutions, washing ways and eluents used in the enrichment of glycopeptides were investigated. When using the ammonium bicarbonate buffer (100 mmol/L, pH 8.0) as the incubation so- lution and incubated for 1 h, the maximum number of enriched glycopeptides from horseradish peroxidase (HRP) digests could be obtained. After enrichment, washing the prepared material with the incubation solution twice for 5 min, nonglyco- peptides interference can be in a large extent removed. The enriched glycopeptides binding to the surface of MCM-41-GLYMO-APB could be released when using 1% trifluoroacetic acid containing 2,5-dihydroxybenzoic acid as the eluent, the operating steps could be simplified when using this optimized eluent because there was no need of adding the MALDI matrix solution afterwards. Therefore, this optimized protocol provided a reference for the glycopeptide enrichment with boronic acid-functionalized materials, and it also laid the foundation for the research of utilizing boronic acid-based chemical method to the glycopeptide enrichment in biological samples. Meanwhile, the highly ordered hexagonal cylindrical mesoporous structure of MCM-41-GLYMO-APB provided the possibility of being used for the research of glycopeptidome in biological samples. Keywords glycoproteins; boronic acid-functionalized mesoporous nanomaterial; enrichment; mass spectrometry