DN承认欧盟FP7项目PEPDIODE（GA 256672）和REGPOT-2012-2013-1（GA 316157），并从以色列科学基金会个人研究资助（编号十分之二百六十八 ）的支持。我们感谢什穆埃尔·鲁宾斯坦教授，工程学院和应用科学，哈佛大学，马萨诸塞州剑桥，美国采取的共聚焦显微镜图像。

1.准备叶绿素a股票方案<ol><li>关键步骤:执行叶绿素准备的所有步骤在化学罩，在绿光（520纳米），或在黑暗中，以尽量减少光损伤。始终冻结颜料存放前添加氮气或氩气。确保所有的溶剂均为分析纯。 </li><li>称取约5mg仅含有叶绿素a在类囊体膜冻干螺旋藻细胞或其它蓝藻细胞，并使用研钵和杵粉碎它。 </li><li>加载破碎细胞到玻璃柱以除去类胡萝卜素用约50-100毫升100％丙酮洗涤。丢弃洗脱橙/绿色部分。 注意:如果橙级分没有与100ml丙酮洗脱继续用丙酮洗涤该细胞直至绿色分数开始洗脱。 </li><li>用100％甲醇交换丙酮并收集含有叶绿素a的绿色部分。洗脱FRACT的体积离子可以50-100毫升之间变化。在开始时，将洗脱级分具有深绿颜色，这在洗脱结束时变成浅绿色。当洗脱馏分的颜色变成浅绿色停止洗脱。 </li><li>使用旋转蒸发器，直到萃取液完全干燥蒸发甲醇。不要使用热到解决方案;确保蒸发器的水浴温度不超过30℃。 注:蒸发的时间取决于甲醇馏分的体积被蒸发上，并且可以为10-60分钟之间变化。它完全干燥的提取物是很重要的。 </li><li>溶解从干燥的提取物中的颜料的乙醚（约5-10毫升）的小体积，并通过脱脂棉过滤。确保颜料​​过滤前完全溶于乙醚。 </li><li>蒸发乙醚，直至颜料完全干燥（10-30分钟）。 注意:干颜料可以与吹扫和氮气或氩气下保持在-20℃下，在黑暗中，直到进一步处理。 </li><li>溶于最小音量可能100％的甲醇（约1毫升）的干颜料，即使不是一切都完全暂停。 4毫升丙酮添加到溶液中，并以完全溶解该色素轻轻拂去玻璃。 </li><li>使用巴斯德吸管轻轻装载样品到DEAE柱在100％丙酮琼脂糖平衡。 </li><li>洗脱类胡萝卜素（橙黄色带）用100％丙酮。然后，洗脱叶绿素a（绿色带）以3:1体积/体积的丙酮/甲醇混合物。 注意:丙酮和丙酮/甲醇混合物的体积是大约相当于DEAE的体积琼脂糖装在柱上。 </li><li>验证叶绿素使用68用薄层色谱纯度 :25:5:2二氯甲烷/正-己烷/异丙醇/甲醇（体积/体积）混合物作为洗脱液17。 </li><li>使用旋转蒸发器蒸发溶剂，直至叶绿素a是完全干燥（10-60分钟）。 注意:干叶绿素a可以用氮气或氩气吹扫，并在黑暗中在-20℃氩气氛下贮存。 </li><li>通过再溶解干燥的叶绿素a在2-4毫升100％乙醇的制备叶绿素原液 。 注:叶绿素a的在663纳米的消光系数是74400厘米-1 M -1（83.3 -1（毫克/毫升）-1）的乙醇。一个典型的库存溶液应具有1860对应于25mM的（23毫克/毫升）的浓度的OD。加入20微升这股到含有5ml有机相，并在1ml的水相的结果1毫克WSCP中的混合物与10倍摩尔过量的叶绿素的对比乳液WSCP。 </li></ol> 2.准备乳液的有机相注意:该乳液的有机相由含有4.5％（体积/体积）跨度80的矿物油，和0.4％（体积/体积）吐温80。 <ol><li>称重在50ml管0.2克的T吐温80，1.8克司盘80的，与38克矿物油。拌匀所有组件和降温冰上。 注意:将有机相可被存储在4℃至一周。 </li></ol> 3.准备乳液的水相注意:该乳液的水相可由任何纯化WSCP，或细菌过度WSCP的粗提取物。 <ol><li>制备含有纯化WSCP水相。 <ol><li>生长含WSCP质粒12在1升的LB培养基中，在37℃，直至0.3-0.6 OD 大肠杆菌 BL21细菌。 </li><li>通过添加1mM的IPTG诱导蛋白质的表达。诱导后细菌生长在30℃，12〜16小时。 </li><li>收获细菌离心5000 xg离心在4℃下10分钟。 </li><li>溶解在冰上的结合缓冲液和超声颗粒（30秒时，15秒关，五次）。使用10个毫升缓冲液从250毫升离心获得的沉淀LB培养基与细胞过度的蛋白质。 注意:根据纯化方法，该结合缓冲液可以由的100mM磷酸缓冲液，pH 7.2或的50mM Tris，pH值7.5，500 mM氯化钠，5mM的EDTA中。 </li><li>降速细胞裂解物在12000×g离心30分钟，在4℃下。 </li><li>通过亲和层析纯化WSCP。取决于融合到WSCP标签，使用适当的可商购的​​亲和层析系统的蛋白质纯化按照生产商的说明纯化重组蛋白。 </li><li>于乳液制剂中，使用在50mM纯化WSCP磷酸盐缓冲液，pH 7.8。确保用于重建的最终蛋白量为0.5-1.0毫克每1ml缓冲液中。 </li></ol></li><li>制备含有与WSCP粗细菌裂解液的水相。 <ol><li>生长含于250毫升LB培养基的质粒表达WSCP在37℃，直到OD 0.3-0.6 大肠杆菌 BL21细胞。 </li><li>诱导与蛋白表达1mM的IPTG，在30℃过夜生长的细菌。 </li><li>收获细菌细胞通过离心5000 xg离心在4℃下10分钟。 </li><li>溶解沉淀在1-2毫升的50mM磷酸钠缓冲液pH 7.8，声处理（在30秒钟，15秒关，三次）和离心机以12,000 xg离心在4℃下30分钟。 </li><li>通过混合0.125毫升上清液与0.875毫升50mM磷酸钠缓冲液pH 7.8的制备乳液的水相。 </li></ol></li></ol> 4. WSCP的大会在乳液叶绿素a <ol><li>转印5毫升油 - 表面活性剂混合物到玻璃小瓶中，并冷却在冰上。移液之前，验证该有机相的所有组分充分混合并有表面活性剂和矿物油之间没有相分离。 </li><li>加入1毫升冰冷的水相的如第3制备至5ml有机相。 </li><li>在9500转在冰上混合使用2分钟组织匀浆两个阶段。</li><li>关键的一步:从这个阶段上，执行以尽量减少光损伤下绿光（520纳米）的所有进一步的步骤。加入20μl的25mM叶绿素的储备液（见1.13），以该乳液。通过轻弹和反相的玻璃小瓶分散。确保叶绿素均匀地分布在乳液中。 </li><li>孵育乳剂在黑暗中在冰上1-2小时。 </li><li>为了打破从有机相中的乳液和单独的水滴转移乳剂1.5毫升的塑料管和离心机在14,000rpm xg离心在室温下5分钟。 注意:如果组件是成功的，下层水相应该具有绿色。 </li><li>配置上油相和加入1 ml矿物油。由涡或彻底翻转管沉淀的乳液拌匀矿物油。降速样品以14,000 xg离心在室温下5分钟。重复此步骤，直到一个明确的半月板分离水和矿工人油相，而没有任何中间的乳液得到的。 </li><li>执行在化学罩此步骤。水性和矿物油相清楚地分离后，除去矿物油和加入1ml水饱和的乙醚。涡旋并在14000 xg离心降速样品在室温下5分钟。重复此步骤两次。 </li><li>第二次离心后，除去乙醚，留下打开引擎盖5-20分钟的管子。 </li><li>最后，装载包含WSCP /叶绿素复合水相到脱盐柱上，用适当的进一步的实验缓冲液洗脱。 注:蛋白质是在宽范围的pH（6.0-7.5）的磷酸盐和Tris缓冲液稳定。样品可以储存在避光长达一个月4℃。 </li></ol>

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作者宣称，他们没有竞争的经济利益。

我们的目标是开发用于与疏水性颜料的水溶性叶绿素结合蛋白的组装一个新的通用系统。这表明，基于W / O乳液的新重组系统是被证明为从球芽甘蓝，花椰菜，日本辣根和弗吉尼亚pepperweed在大肠杆菌中重组表达WSCP脱辅基蛋白的组装工作的一般方法大肠杆菌 。这里的结果是从1毫克WSCP的重构带有10倍摩尔过量的叶绿素a的。然而也可以使用较低浓度WSCP用于reconstitutions和不同的Chl /...

疏水性颜料如叶绿素（的Chls），菌绿素（BChls）和类胡萝卜素在光合作用反应中心，捕光蛋白是进行电子传输和光能量采集和传输的主要辅助因子。反应中心和大部分叶绿素结合光收获物是跨膜蛋白。非氧光合绿硫细菌1,2的Fenna -马修斯-奥尔森（FMO）的蛋白质，和甲藻3的甲藻素-叶绿素蛋白（PCP）是水溶性的捕光蛋白的特殊例子。水溶性叶绿素结合蛋白十字花科 ， 蓼 ， 藜科 （WSCPs）和苋植物4,5另一个独特的例子，但在对比FMO和PCP，这些既不参与捕光也不在任何主光合反应的和它们的精确的physiological功能还不清楚5-8。它们的高叶绿素结合亲和力，导致一个建议的功能的Chls瞬态载体和叶绿素衍生物9,10。另外，有人推测WSCP起着受损细胞清除的Chls作用和防止叶绿素引起的光氧化损伤7,11-13。最近，有人提出WSCP用作蛋白酶抑制剂和播放食草动物性过程中的作用，以及花的发育过程中14调节细胞死亡。 WSCPs根据其光物理性质分为两大类。第一类（I类， 如 :从藜 ）可根据照明进行光转化。类从芸苔属植物，即不发生光转化5,10-二WSCPs，被进一步细分为IIa类（ 例如 ， 甘蓝，萝卜 ）和Ⅱb族（ 例如 ，来自北美独行菜 </eM&gt;）。 IIb类WSCP从北美独行菜的结构用X射线晶体学的2.0 A分辨率8解决。它揭示了一个对称的同源四聚体，其中所述蛋白质亚基形成疏水核心。每个亚基结合的单个叶绿素，这导致在四个紧密堆积的Chls的简单所有叶绿素布置使得WSCPs潜在有用的模型系统用于研究叶绿素 - 蛋白复合物的结合和组装core.This内紧密排列，且相邻的Chls的影响和对个人的Chls的光谱和电子特性蛋白的环境中。此外，其可提供用于构建可用于人工光合作用设备捕光模块人工叶绿素结合蛋白的模板。 天然WSCPs的严谨的研究是不可行的，因为从植物中纯化复合物总是含有与叶绿素a的不同组合四聚体的异质混合物和叶绿素b 9。因此，需要一种用于组装具有的Chls 体外重组表达WSCPs的方法。这是通过的Chls的可忽略的水溶解度，这使得它不可能通过简单地与在水溶液中的颜料混合水溶性脱辅基蛋白聚集在体外复通过混合15被证明与类囊体膜的脱辅基蛋白的挑战。 在体外组装，但这种方法仅限于原生的Chls存在于类囊体的。 Schmidt 等人 。报告了在大肠杆菌中重组表达组氨酸标记的蛋白质组装与来自花椰菜（CaWSCP）WSCP几个Chl和Bchl衍生物大肠杆菌它固定到一个镍亲和柱和引进洗涤剂溶解11叶绿素衍生物。成功复溶从A重组WSCPs的拟南芥 6，和球芽甘蓝（BoWSCP），日野萝卜（RshWSCP）的还报道ð弗吉尼亚pepperweed（LvWSCP）类似的方法由被。 在这里，我们提出了一个新颖，装配与WSCP的Chls不需要标记或固定蛋白质一般情况下，直接的方法。它依赖于从在矿物油中的水溶性脱辅基蛋白的它们的水溶液制备乳液。因此蛋白质被封装在水包油（W / O）具有非常高的表面微滴体积比16。然后疏水辅因子被溶解在油中并且容易引入到从油相的液滴。我们使用方法报到WSCP脱辅基蛋白的几个变体在大肠杆菌重组表达的组装大肠杆菌与叶绿素a。我们证明从中可以用作用于开发新颖叶绿素结合蛋白筛选系统WSCP过表达的细菌粗裂解物的装配。

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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Mineral oil</td>
			<td style="width:83px;">Sigma</td>
			<td style="width:119px;">M5904</td>
			<td style="width:196px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Span80</td>
			<td style="width:83px;">Sigma</td>
			<td style="width:119px;">85548</td>
			<td style="width:196px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tween80</td>
			<td style="width:83px;">Sigma</td>
			<td style="width:119px;">P8074</td>
			<td style="width:196px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Bio-Scale Mini Profinity eXact Cartridges</td>
			<td style="width:83px;">Bio Rad</td>
			<td style="width:119px;">10011164</td>
			<td style="width:196px;">Affinity chromatography for WSCP purification with native sequence.</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">His Trap HF column</td>
			<td style="width:83px;">GE Healthcare Life Science</td>
			<td style="width:119px;">17-5248-02</td>
			<td style="width:196px;">Affinity chromatography for WSCP purification with His-tag</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">DEAE Sepharose Fast Flow</td>
			<td style="width:83px;">GE Healthcare Life Science</td>
			<td style="width:119px;">17-0709-01</td>
			<td style="width:196px;">Chromatography medium for chlorophyll purification</td>
		</tr>
	</tbody>
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water in oil emulsions: a new system for assembling water-soluble chlorophyll-binding proteins with hydrophobic pigments

叶绿素（的Chls）和菌绿素（BChls）是进行光合作用的光捕获和电子传递的主要辅助因子。其功能主要取决于他们的大而复杂的多亚基跨膜蛋白复合体中的特定组织。为了在分子水平上，这些配合物如何促进太阳能转换了解，重要的是要了解蛋白质颜料，和颜料颜料的相互作用，以及它们对兴奋动力学效果。获得这样的理解的一种方式是通过构建和学习的Chls的配合物与简单的水溶性的重组蛋白。然而，掺入亲脂性的Chls和BChls成水溶性蛋白是困难的。此外，不存在一般的方法，它可以用于与疏水性颜料的水溶性蛋白质的组装。在这里，我们证明根据水包油乳剂的简单和高通量系统，能使屁股疏水的Chls水溶性蛋白质的embly。的新方法，通过组装与叶绿素a 十字花科植物（WSCP）的水溶性叶绿素结合蛋白的重组形式的验证。我们证明叶绿素的成功组装表达E. WSCP 的使用原油裂解大肠杆菌细胞，其可以用于开发遗传筛选系统，用于新的水溶性叶绿素结合蛋白，以及叶绿素-蛋白相互作用和装配过程的研究。

重组WSCP载脂蛋白组装与根据上一节中描述的方案的W / O乳剂叶绿素a。该协议使用含有任一纯WSCPs，或裂解物的大肠杆菌细胞过度WSCP（ 图1）的水相来实现。该协议是简单，快速，不需要任何特殊的设备，除了组织匀浆。 吸光度和叶绿素的CD谱四个重组WSCP变体，即RshWSCP，CaWSCP，BoWSCP和LvWSCP 一个?...

Watch this Scientific Journal Video about Water in Oil Emulsions: A New System for Assembling Water-soluble Chlorophyll-binding Proteins with Hydrophobic Pigments at JoVE.com

油包水乳液：用疏水颜料组装水溶性叶绿素结合蛋白的新系统化学|视频

Water in Oil Emulsions: A New System for Assembling Water-soluble Chlorophyll-binding Proteins with Hydrophobic Pigments

该原稿描述用于装配与基于水包油乳液的疏水颜料的水溶性蛋白质的简单和高通量方法。我们证明该方法在本地叶绿素大会的有效性与重组水溶性叶绿素的四种变体结合蛋白（WSCPs）的芸苔属植物在大肠杆菌中表达大肠杆菌 。

水在油乳剂:与疏水性颜料装配水溶性叶绿素结合蛋白的新系统

Chlorophylls (Chls) and bacteriochlorophylls (BChls) are the primary cofactors that carry out photosynthetic light harvesting and electron transport. ...

water-oil-emulsions-new-system-for-assembling-water-soluble

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14.5K 次观看.  Weizmann Institute of Science. 该原稿描述用于装配与基于水包油乳液的疏水颜料的水溶性蛋白质的简单和高通量方法。我们证明该方法在本地叶绿素大会的有效性与重组水溶性叶绿素的四种变体结合蛋白（WSCPs）的芸苔属植物在大肠杆菌中表达大肠杆菌 。 

视频: Water in Oil Emulsions: A New System for Assembling Water-soluble Chlorophyll-binding Proteins with Hydrophobic Pigments

High-pressure Sapphire Cell for Phase Equilibria Measurements of CO2/Organic/Water Systems

The high pressure sapphire cell apparatus was constructed to visually determine the composition of multiphase systems without physical sampling. Specifically, the sapphire cell enables visual data collection from multiple loadings to solve a set of material balances to precisely determine phase composition. Ternary phase diagrams can then be established to determine the proportion of each component in each phase at a given condition. In principle, any ternary system can be studied although ternary systems (gas-liquid-liquid) are the specific examples discussed herein. For instance, the ternary THF-Water-CO2 system was studied at 25 and 40&#160;&#176;C and is described herein. Of key importance, this technique does not require sampling. Circumventing the possible disturbance of the system equilibrium upon sampling, inherent measurement errors, and technical difficulties of physically sampling under pressure is a significant benefit of this technique. Perhaps as important, the sapphire cell also enables the direct visual observation of the phase behavior. In fact, as the CO2 pressure is increased, the homogeneous THF-Water solution phase splits at about 2 MPa. With this technique, it was possible to easily and clearly observe the cloud point and determine the composition of the newly formed phases as a function of pressure.
The data acquired with the sapphire cell technique can be used for many applications. In our case, we measured swelling and composition for tunable solvents, like gas-expanded liquids, gas-expanded ionic liquids and Organic Aqueous Tunable Systems (OATS)1-4. For the latest system, OATS, the high-pressure sapphire cell enabled the study of (1) phase behavior as a function of pressure and temperature, (2) composition of each phase (gas-liquid-liquid) as a function of pressure and temperature and (3) catalyst partitioning in the two liquid phases as a function of pressure and composition. Finally, the sapphire cell is an especially effective tool to gather accurate and reproducible measurements in a timely fashion.

High-pressure Sapphire Cell for Phase Equilibria Measurements of CO2/Organic/Water Systems

The high pressure sapphire cell apparatus is a unique tool to study, without sampling, phase behavior under a wide range of pressures. Using a cathetometer, very precise volume measurements can be recorded to measure liquid expansion and phase composition. Thus, this synthetic method enables the study of (1) phase equilibria of multi-component mixtures and (2) the partition behavior of catalyst or model compounds as a function of pressure.

高压蓝宝石细胞对CO的相平衡测量<sub&gt; 2</sub&gt; /有机/水系统

The high pressure sapphire cell apparatus was constructed to visually determine the composition of multiphase systems without physical sampling. ...

科研

化学

In Situ Characterization of Hydrated Proteins in Water by SALVI and ToF-SIMS

This work demonstrates in situ characterization of protein biomolecules in the aqueous solution using the System for Analysis at the Liquid Vacuum Interface (SALVI) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). The fibronectin protein film was immobilized on the silicon nitride (SiN) membrane that forms the SALVI detection area. During ToF-SIMS analysis, three modes of analysis were conducted including high spatial resolution mass spectrometry, two-dimensional (2D) imaging, and depth profiling. Mass spectra were acquired in both positive and negative modes. Deionized water was also analyzed as a reference sample. Our results show that the fibronectin film in water has more distinct and stronger water cluster peaks compared to water alone. Characteristic peaks of amino acid fragments are also observable in the hydrated protein ToF-SIMS spectra. These results illustrate that protein molecule adsorption on a surface can be studied dynamically using SALVI and ToF-SIMS in the liquid environment for the first time.

In Situ Characterization of Hydrated Proteins in Water by SALVI and ToF-SIMS

This work presents a protocol for liquid handling and sample introduction to a microchannel for in situ time-of-flight secondary ion mass spectrometry analysis of protein biomolecules in an aqueous solution.

在水中水合蛋白原位表征SALVI和TOF-SIMS

This work demonstrates in situ characterization of protein biomolecules in the aqueous solution using the System for Analysis at the Liquid Vacuum ...

Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry

The chemical linking or bioconjugation of proteins to fluorescent dyes, drugs, polymers and other proteins has a broad range of applications, such as the development of antibody drug conjugates (ADCs) and nanomedicine, fluorescent microscopy and systems chemistry. For many of these applications, specificity of the bioconjugation method used is of prime concern. The Michael addition of maleimides with cysteine(s) on the target proteins is highly selective and proceeds rapidly under mild conditions, making it one of the most popular methods for protein bioconjugation.
We demonstrate here the modification of the only surface-accessible cysteine residue on yeast cytochrome c with a ruthenium(II) bisterpyridine maleimide. The protein bioconjugation is verified by gel electrophoresis and purified by aqueous-based fast protein liquid chromatography in 27% yield of isolated protein material. Structural characterization with MALDI-TOF MS and UV-Vis is then used to verify that the bioconjugation is successful. The protocol shown here is easily applicable to other cysteine - maleimide coupling of proteins to other proteins, dyes, drugs or polymers.

Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry

This protocol details the important steps required for the bioconjugation of a cysteine containing protein to a maleimide, including reagent purification, reaction conditions, bioconjugate purification and bioconjugate characterization.

蛋白质生物缀合物的合成<em&gt;通过</em&gt;半胱氨酸马来酰亚胺化学

The chemical linking or bioconjugation of proteins to fluorescent dyes, drugs, polymers and other proteins has a broad range of applications, such as the ...

Synthesis of a Water-soluble Metal&#8211;Organic Complex Array

We demonstrate a method for the synthesis of a water-soluble multimetallic peptidic array containing a predetermined sequence of metal centers such as Ru(II), Pt(II), and Rh(III). The compound, named as a water-soluble metal-organic complex array (WSMOCA), is obtained through 1) the conventional solution-chemistry-based preparation of the corresponding metal complex monomers having a 9-fluorenylmethyloxycarbonyl (Fmoc)-protected amino acid moiety and 2) their sequential coupling together with other water-soluble organic building units on the surface-functionalized polymeric resin by following the procedures originally developed for the solid-phase synthesis of polypeptides, with proper modifications. Traces of reactions determined by mass spectrometric analysis at the representative coupling steps in stage 2 confirm the selective construction of a predetermined sequence of metal centers along with the peptide backbone. The WSMOCA cleaved from the resin at the end of stage 2 has a certain level of solubility in aqueous media dependent on the pH value and/or salt content, which is useful for the purification of the compound.

A potential general method for the synthesis of water-soluble multimetallic peptidic arrays containing a predetermined sequence of metal centers is presented.

一个水溶性金属 - 有机络合物阵列合成

We demonstrate a method for the synthesis of a water-soluble multimetallic peptidic array containing a predetermined sequence of metal centers such as ...

NMR Spectroscopy as a Robust Tool for the Rapid Evaluation of the Lipid Profile of Fish Oil Supplements

The western diet is poor in n-3 fatty acids, therefore the consumption of fish oil supplements is recommended to increase the intake of these essential nutrients. The objective of this work is to demonstrate the qualitative and quantitative analysis of encapsulated fish oil supplements using high-resolution 1H and 13C NMR spectroscopy utilizing two different NMR instruments; a 500 MHz and an 850 MHz instrument. Both proton (1H) and carbon (13C) NMR spectra can be used for the quantitative determination of the major constituents of fish oil supplements. Quantification of the lipids in fish oil supplements is achieved through integration of the appropriate NMR signals in the relevant 1D spectra. Results obtained by 1H and 13C NMR are in good agreement with each other, despite the difference in resolution and sensitivity between the two nuclei and the two instruments. 1H NMR offers a more rapid analysis compared to 13C NMR, as the spectrum can be recorded in less than 1 min, in contrast to 13C NMR analysis, which lasts from 10 min to one hour. The 13C NMR spectrum, however, is much more informative. It can provide quantitative data for a greater number of individual fatty acids and can be used for determining the positional distribution of fatty acids on the glycerol backbone. Both nuclei can provide quantitative information in just one experiment without the need of purification or separation steps. The strength of the magnetic field mostly affects the 1H NMR spectra due to its lower resolution with respect to 13C NMR, however, even lower cost NMR instruments can be efficiently applied as a standard method by the food industry and quality control laboratories.

Here, high-resolution 1H and 13C Nuclear Magnetic Resonance (NMR) spectroscopy was used as a rapid and reliable tool for quantitative and qualitative analysis of encapsulated fish oil supplements.

核磁共振作为鱼油补充剂的血脂的快速评价一个强大的工具

The western diet is poor in n-3 fatty acids, therefore the consumption of fish oil supplements is recommended to increase the intake of these essential ...

Probing the Structure and Dynamics of Interfacial Water with Scanning Tunneling Microscopy and Spectroscopy

Water/solid interfaces are ubiquitous and play a key role in many environmental, biophysical, and technological processes. Resolving the internal structure and probing the hydrogen-bond (H-bond) dynamics of the water molecules adsorbed on solid surfaces are fundamental issues of water science, which remains a great challenge owing to the light mass and small size of hydrogen. Scanning tunneling microscopy (STM) is a promising tool for attacking these problems, thanks to its capabilities of sub-&#197;ngstr&#246;m spatial resolution, single-bond vibrational sensitivity, and atomic/molecular manipulation. The designed experimental system consists of a Cl-terminated tip and a sample fabricated by dosing water molecules in situ onto the Au(111)-supported NaCl(001) surfaces. The insulating NaCl films electronically decouple the water from the metal substrates, so the intrinsic frontier orbitals of water molecules are preserved. The Cl-tip facilitates the manipulation of the single water molecules, as well as gating the orbitals of water to the proximity of Fermi level (EF) via tip-water coupling. This paper outlines the detailed methods of submolecular resolution imaging, molecular/atomic manipulation, and single-bond vibrational spectroscopy of interfacial water. These studies open up a new route for investigating the H-bonded systems at the atomic scale.

Here, we present a protocol to investigate the structure and dynamics of interfacial water at the atomic scale, in terms of submolecular resolution imaging, molecular manipulation, and single-bond vibrational spectroscopy.

用扫描隧穿显微镜和光谱法探讨界面水的结构和动力学

Water/solid interfaces are ubiquitous and play a key role in many environmental, biophysical, and technological processes. Resolving the internal ...

Heterogeneous Removal of Water-Soluble Ruthenium Olefin Metathesis Catalyst from Aqueous Media Via Host-Guest Interaction

A highly efficient transition metal catalyst removal method is developed. The water-soluble catalyst contains a newly-designed NHC ligand for the catalyst removal via host-guest interactions. The new NHC ligand possesses an adamantyl (guest) tethered linear ethylene glycol units for hydrophobic inclusion into the cavity of a &#946;-cyclodextrin (&#946;-CD) host compound. The new NHC ligand was applied to a Ru-based olefin metathesis catalyst. The Ru catalyst demonstrated excellent activity in representative ring-closing metathesis (RCM) and ring-opening metathesis polymerization (ROMP) reactions in aqueous media as well as organic solvent, CH2Cl2. After the reaction was complete, the lingering Ru residue was removed from the aqueous solution with the efficiency of more than 99% (53 ppm of Ru residue) by simple filtration utilizing a host-guest interaction between insoluble silica-grafted &#946;-CD (host) and the adamantyl moiety (guest) on the catalyst. The new Ru catalyst also demonstrated high removal efficiency via extraction when the reaction is run in organic solvent by partitioning the crude reaction mixture between layers of diethyl ether and water. In this way, the catalyst stays in aqueous layer only. In organic layer, the residual Ru amount was only 0.14 ppm in the RCM reactions of diallyl compounds.

A removable water-soluble N-heterocyclic carbene (NHC) ligand in aqueous media via host-guest interaction has been developed. We demonstrated representative olefin metathesis reactions in water as well as in dichloromethane. Via either host-guest interaction or extraction, the residual ruthenium (Ru) catalyst was as low as 0.14 ppm after the reaction.

水介质中水溶性钌烯烃置换催化剂的异质化去除与寄主交互作用

A highly efficient transition metal catalyst removal method is developed. The water-soluble catalyst contains a newly-designed NHC ligand for the catalyst ...

Stereolithographic 3D Printing with Renewable Acrylates

The accessibility of cost-competitive renewable materials and their application in additive manufacturing is essential for an efficient biobased economy. We demonstrate the rapid prototyping of sustainable resins using a stereolithographic 3D printer. Resin formulation takes place by straightforward mixing of biobased acrylate monomers and oligomers with a photoinitiatior and optical absorber. Resin viscosity is controlled by the monomer to oligomer ratio and is determined as a function of shear rate by a rheometer with parallel plate geometry. A stereolithographic apparatus charged with the biobased resins is employed to produce complex shaped prototypes with high accuracy. The products require a post-treatment, including alcohol rinsing and UV irradiation, to ensure complete curing. The high feature resolution and excellent surface finishing of the prototypes is revealed by scanning electron microscopy.

A protocol for additive manufacturing with renewable photopolymer resins on a stereolithography apparatus is presented.

Stereolithographic 3D 印刷用可再生丙烯酸酯

The accessibility of cost-competitive renewable materials and their application in additive manufacturing is essential for an efficient biobased economy. ...

A New Straightforward Method for Lipophilicity (logP) Measurement using 19F NMR Spectroscopy

Fluorination has become an effective tool to optimize physicochemical properties of bioactive compounds. One of the applications of fluorine introduction is to modulate the lipophilicity of the compound. In our group, we are interested in the study of the impact of fluorination on lipophilicity of aliphatic fluorohydrins and fluorinated carbohydrates. These are not UV-active, resulting in a challenging lipophilicity determination. Here, we present a straightforward method for the measurement of lipophilicity of fluorinated compounds by 19F NMR spectroscopy. This method requires no UV-activity. Accurate solute mass, solvent and aliquot volume are also not required to be measured. Using this method, we measured the lipophilicities of a large number of fluorinated alkanols and carbohydrates.

A New Straightforward Method for Lipophilicity logP Measurement using 19F NMR Spectroscopy

A novel and straightforward variation of the shake-flask method was developed for accurate lipophilicity measurement of fluorinated compounds by 19F NMR spectroscopy.

一种利用19f 核磁共振光谱测定脂质性 (日志 p) 的新方法

Fluorination has become an effective tool to optimize physicochemical properties of bioactive compounds. One of the applications of fluorine introduction ...

Synthesis and Bioconjugation of Thiol-Reactive Reagents for the Creation of Site-Selectively Modified Immunoconjugates

Maleimide-bearing bifunctional probes have been employed for decades for the site-selective modification of thiols in biomolecules, especially antibodies. Yet maleimide-based conjugates display limited stability in vivo because the succinimidyl thioether linkage can undergo a retro-Michael reaction. This, of course, can lead to the release of the radioactive payload or its exchange with thiol-bearing biomolecules in circulation. Both of these processes can produce elevated activity concentrations in healthy organs as well as decreased activity concentrations in target tissues, resulting in reduced imaging contrast and lower therapeutic ratios. In 2018, we reported the creation of a modular, stable, and easily accessible phenyloxadiazolyl methyl sulfone reagent &#8212; dubbed &#8216;PODS&#8217; &#8212; as a platform for thiol-based bioconjugations. We have clearly demonstrated that PODS-based site-selective bioconjugations reproducibly and robustly create homogenous, well-defined, highly immunoreactive, and highly stable radioimmunoconjugates. Furthermore, preclinical experiments in murine models of colorectal cancer have shown that these site-selectively labeled radioimmunoconjugates exhibit far superior in vivo performance compared to radiolabeled antibodies synthesized via maleimide-based conjugations. In this protocol, we will describe the four-step synthesis of PODS, the creation of a bifunctional PODS-bearing variant of the ubiquitous chelator DOTA (PODS-DOTA), and the conjugation of PODS-DOTA to the HER2-targeting antibody trastuzumab.

In this protocol, we will describe the synthesis of PODS, a phenyoxadiazolyl methyl sulfone-based reagent for the site-selective attachment of cargos to the thiols of biomolecules, particularly antibodies. In addition, we will describe the synthesis and characterization of a PODS-bearing bifunctional chelator and its conjugation to a model antibody.

硫醇反应试剂的合成与生物共轭作用--用于原位选择性修饰免疫共轭的生成

Maleimide-bearing bifunctional probes have been employed for decades for the site-selective modification of thiols in biomolecules, especially antibodies. ...

水在油乳剂:与疏水性颜料装配水溶性叶绿素结合蛋白的新系统

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