Acid cleavable surface enhanced raman tagging for protein detection.
Acid cleavable surface enhanced raman tagging for protein detection.
复制标题
用于蛋白质检测的酸可裂解表面增强拉曼标记。
DOI：
10.1039/c0an00708k
复制
发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Sygula,Andrzej
中科院分区：
文献类型：
--
作者：
Zhang,Dongmao;Vangala,Karthikeshwar;Li,Shaoyong;Yanney,Michael;Xia,Hao;Zou,Sige;Sygula,Andrzej
染料 共轭是提高生物分子的表面增强拉曼检测灵敏度的常用策略。报道了一种新的表面增强拉曼光谱标记策略的概念验证研究，称为酸可裂解Sers标签（ACST）方法。使用罗丹明B作为起始材料，我们制备了第一个ACST原型，其由从远端开始的Sers标签部分（STM）、酸可切割的接头和蛋白质反应部分组成。ACST标签的完全酸裂解是在非常温和的条件下实现的，该条件是在室温下的1.5%三氟乙酸（TFA）水溶液。使用牛血清白蛋白（BSA）作为模型蛋白，证明了这种ACST标记的蛋白质的Sers检测。虽然完整的ACST-BSA的Sers光谱完全由STM的荧光信号主导，但用酸裂解的ACST-BSA缀合物可以容易地获得高质量的Sers光谱。从蛋白质中分离酸裂解的STM进一步增强了Sers灵敏度。用酸裂解的ACST-BSA缀合物实现的当前Sers检测灵敏度在BSA浓度方面为1.5 nM，在ACST含量方面为1.5 nM。裂解ACST-BSA偶联物的动态范围跨越4个数量级，蛋白质浓度为10 nM至100 μM。通过共振拉曼采集可以实现Sers灵敏度的进一步提高。这种可切割的标记策略也可用于消除基于荧光的生物分子检测中的蛋白质干扰。
Dye conjugation is a common strategy improving the surface enhanced Raman detection sensitivity of biomolecules. Reported is a proof-of-concept study of a novel surface enhanced Raman spectroscopic tagging strategy termed as acid-cleavable SERS tag (ACST) method. Using Rhodamine B as the starting material, we prepared the first ACST prototype that consisted of, from the distal end, a SERS tag moiety (STM), an acid-cleavable linker, and a protein reactive moiety. Complete acid cleavage of the ACST tags was achieved at a very mild condition that is 1.5% trifluoroacetic acid (TFA) aqueous solution at room temperature. SERS detection of this ACST tagged protein was demonstrated using bovine serum albumin (BSA) as the model protein. While the SERS spectrum of intact ACST-BSA was entirely dominated by the fluorescent signal of STM, quality SERS spectra can be readily obtained with the acid cleaved ACST-BSA conjugates. Separation of the acid cleaved STM from protein further enhances the SERS sensitivity. Current SERS detection sensitivity, achieved with the acid cleaved ACST-BSA conjugate is ∼5 nM in terms of the BSA concentration and ∼1.5 nM in ACST content. The dynamic range of the cleaved ACST-BSA conjugate spans four orders of magnitudes from ∼10 nM to ∼100 μM in protein concentrations. Further improvement in the SERS sensitivity can be achieved with resonance Raman acquisition. This cleavable tagging strategy may also be used for elimination of protein interference in fluorescence based biomolecule detection.