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ZnCl₂ 降水辅助样本准备用于蛋白质组分析

Qiqing He^1,2,3, Qingjing Chen^2,3,4, Dongxue Wang^2,3,5

¹Department of Chemistry, School of Science, Southern University of Science and Technology, Shenzhen, China.

|July 31, 2025

概括

Cl2沉辅助样本制备 (ZASP) 提供了一种具有成本效益和高效的蛋白质组分析方法. 该方案有效地去除杂质,并回收超过90%的蛋白质,优于其他样品制备技术.

科学领域:

蛋白质组学和质谱学
生物化学和分子生物学

背景情况:

传统的蛋白质组样本制备方法往往难以去除洗剂和杂质,影响蛋白质回收和分析灵敏度.
硫酸二酸 (SDS) 和尿素等强化学物质在生物溶解酸中很常见,但会干扰下游的酶消化和质谱.
开发强大,具有成本效益和高效的样本准备协议对于推进蛋白质组研究至关重要.

研究的目的:

引入和验证用于增强蛋白质组分析的ZnCl2沉辅助样本制备 (ZASP) 协议.
证明ZASP在去除常见污染物和改善蛋白质回收方面的有效性.
为了比较ZASP的性能与已建立的样本准备方法.

主要方法:

使用含有ZnCl2和甲醇的ZASP沉缓冲器 (ZPB) 沉蛋白质,以去除洗剂和杂质.
蛋白质回收率是通过化酸盐与ZPB进行评估的.
用Exploris 480质谱仪使用数据依赖采集 (DDA) 进行蛋白质组分析.
对ZASP的性能进行了基准测试,与基于脱氧化 (SDC) 的消化,乙沉,过辅助样本制备 (FASP) 和单,固相增强样本制备 (SP3) 相比.

主要成果:

在室温下,ZASP在10分钟内从酸盐中恢复了90%以上的蛋白质.
对1μg小鼠小肠蛋白质的分析产生了4,037种蛋白质和25,626种,对于增加输入的产量更高.
在蛋白质和的识别方面,ZASP的表现优于其他方法,获得了4,456种蛋白质和29,871种,缺失裂解率较低 (16.3%) 和高可重复性 (Pearson相关系数为0.96).
对于ZASP,每个样本的成本不到30元人民币,包括素.

结论:

ZASP是一种用户友好,高效,灵敏和具有成本效益的蛋白质样本制备方法,适合标准生物化学实验室.
该协议可以从多种生物样本 (包括细胞和组织 (OCT嵌入式和FFPE) 中进行高蛋白回收和公正分析.
与传统方法相比,ZASP显著提高了蛋白质组数据质量和可重复性.

关键词:

与LCMS不兼容的试剂蛋白质的恢复蛋白质的恢复蛋白质组学. 蛋白质组学.样品的准备样品的准备 ZnCl2诱导蛋白质沉导致的蛋白质沉.

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