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利用原子力显微镜(AFM)测定不利于 SPR 或 QCM 的材料表面的肽-表面吸附自由能。

Determination of peptide-surface adsorption free energy for material surfaces not conducive to SPR or QCM using AFM.

机构信息

Department of Bioengineering, Clemson University, Clemson, South Carolina 29634, United States.

出版信息

Langmuir. 2012 Apr 3;28(13):5687-94. doi: 10.1021/la300315r. Epub 2012 Mar 20.

Abstract

The interactions between peptides and proteins with material surfaces are of primary importance in many areas of biotechnology. While surface plasmon resonance spectroscopy (SPR) and quartz crystal microbalance (QCM) methods have proven to be very useful in measuring fundamental properties characterizing adsorption behavior, such as the free energy of adsorption for peptide-surface interactions, these methods are largely restricted to use for materials that can readily form nanoscale-thick films over the respective sensor surfaces. Many materials including most polymers, ceramics, and inorganic glasses, however, are not readily suitable for use with SPR or QCM methods. To overcome these limitations, we recently showed that desorption forces (F(des)) obtained using a standardized AFM method linearly correlate to standard-state adsorption free energy values (ΔG°(ads)) measured from SPR in phosphate buffered saline (PBS: phosphate buffered 140 mM NaCl, pH 7.4). This approach thus provides a means to determine ΔG°(ads) for peptide adsorption using AFM that can be applied to any flat material surface. In this present study, we investigated the F(des)-ΔG°(ads) correlation between AFM and SPR data in PBS for a much broader range of systems including eight different types of peptides on a set of eight different alkanethiol self-assembled monolayer (SAM) surfaces. The resulting correlation was then used to estimate ΔG°(ads) from F(des) determined by AFM for selected bulk polymer and glass/ceramic materials such as poly(methyl methacrylate) (PMMA), high-density polyethylene (HDPE), fused silica glass, and a quartz (100) surface. The results of these studies support our previous findings regarding the strong correlation between F(des) measured by AFM and ΔG°(ads) determined by SPR, and provides a means to estimate ΔG°(ads) for peptide adsorption on macroscopically thick samples of materials that are not conducive for use with SPR or QCM.

摘要

肽和蛋白质与材料表面的相互作用在许多生物技术领域都具有重要意义。尽管表面等离子体共振光谱(SPR)和石英晶体微天平(QCM)方法已被证明非常适用于测量描述吸附行为的基本特性,例如肽-表面相互作用的吸附自由能,但这些方法在很大程度上仅限于可在各自传感器表面上形成纳米级厚膜的材料。然而,许多材料,包括大多数聚合物、陶瓷和无机玻璃,并不适合使用 SPR 或 QCM 方法。为了克服这些限制,我们最近表明,使用标准化原子力显微镜(AFM)方法获得的解吸力(F(des))与从 SPR 在磷酸盐缓冲盐水(PBS:磷酸盐缓冲 140 mM NaCl,pH 7.4)中测量的标准态吸附自由能值(ΔG°(ads))呈线性相关。这种方法为使用 AFM 确定肽吸附的ΔG°(ads)提供了一种手段,可应用于任何平坦的材料表面。在本研究中,我们研究了 PBS 中 AFM 和 SPR 数据之间的 F(des)-ΔG°(ads)相关性,涵盖了包括八种不同类型的肽在八套不同的烷硫醇自组装单层(SAM)表面上的更广泛的系统。然后,将该相关性用于从 AFM 确定的 F(des)估计选定的大块聚合物和玻璃/陶瓷材料(如聚甲基丙烯酸甲酯(PMMA)、高密度聚乙烯(HDPE)、熔融石英玻璃和石英(100)表面)的ΔG°(ads)。这些研究的结果支持我们之前关于 AFM 测量的 F(des)与 SPR 确定的ΔG°(ads)之间强相关性的发现,并为估计在不适合使用 SPR 或 QCM 的宏观厚样品上的肽吸附的ΔG°(ads)提供了一种手段。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1a56/3319500/a0063b0a0b5d/nihms362747f1.jpg

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