An optical surface plasmon resonance biosensor for determination of tetanus toxin

Surface plasmon resonance (SPR) has been successfully applied for the simple, rapid, and label-free assay of various biomolecules. This assay evaluates a novel wavelength modulation SPR biosensor for the detection of tetanus toxin. The wavelength modulation SPR biosensor is designed based on fixing the incident angle of light and measuring the reflected intensities in the resonance wavelength range spanning 400-800 nm simultaneously. Tetanus toxin (TeNT), one of the most potent toxins known, is synthesized as a 150 kDa single polypeptide chain. The SPR biosensor has been shown to be capable of directly detecting concentration of tetanus toxin as low as 0.028 Lf ml⁻¹. Under selected experimental conditions, the SPR biosensor has a good reproducibility, sensitivity and reversibility. The results illustrate how wavelength modulation SPR biosensor can be used to detect biomolecular interactions.     

Saccharide-branched Cyclodextrins as Targeting Drug Carriers

A synthetic series of heptakis-galactose-branched cyclodextrins (termed CDs) having a longer spacer arm using two amino-caproic acids as an enlarging unit were prepared. Starting with heptakis-amino-β-CD or heptakis-amino-caproic-amide-β-CD, treated with galactosyl-glucono-amide-caproic acid, the new compounds heptakis (Gal-cap1)-CD (4) or heptakis (Gal-cap2)-CD (5) were obtained. The longer galactose spacer arm extremely favors the PNA association. The effect of branch length on K with PNA was enhanced up to 138-fold 3 as well as with DXR enhanced up to 81-fold. Hexakis (Gal-cap2)-CD (6) was prepared and the association constants with rat liver cells were observed to be 2.5 × 10¹⁰ M⁻¹. A multi-high mannose type oligosaccharide branched CD (7) showed a large association constant with DXR up to 1.1 × 10⁹ M⁻¹. The two-dimensional map for the association constants of newly synthesized oligosaccharide-branched CDs toward lectin or liver cells versus the association constants toward a drug (doxorubicin) suggested a method of finding a better targeting drug carrier. The structural effect of the oligosaccharide-CDs showed that the number and length of the branch were dominant factors in designing for enhanced dual recognition.     

Sequential-dissociation kinetics of non-covalent complexes of DNA with multiple proteins in separation-based approach: General theory and its application

Binding of multiple proteins to DNA is crucial in many regulatory cellular processes. The kinetics of assembly and disassembly of DNA–multiple protein complexes is very difficult to study in detail due to the lack of suitable experimental approaches. A separation-based approach has been recently proposed to resolve disassembly kinetics of such complexes. While conceptually simple, the separation-based approach generates experimental data with very complex patterns. The analysis of these patterns is a challenging problem on its own. Here we report on a mathematical approach that can extract a solution for the experimental data obtained in separation-based analysis of sequential dissociation of a DNA complex with multiple proteins. This case describes the dissociation of proteins one-by-one from the complex. Generally speaking, a mathematical solution of such problems requires calculations of multiple integrals. Our approach reduces this procedure to taking double integrals and constructing their superposition. We tested this approach with the experimental data obtained for three-step sequential dissociation of complexes of DNA with two protein copies.     

纳米金标记抗体增强SPR检测大肠杆菌O157∶H7

将金纳米粒子(AuNPs)标记的大肠杆菌O157∶H7(E.coli O157∶H7)的多克隆抗体(PAb)作为二抗,采用氨基偶联法将PAb固定在传感器表面作为一抗,通过三明治方法用双通道表面等离子体子共振(SPR)传感器对E.coli O157∶H7进行检测,并与SPR直接法检测进行了比较.结果表明,直接法的检出限为103cfu/mL,线性范围为103～109cfu/mL;AuNPs增强三明治法的检出限为10 cfu/mL,线性范围为10～1010cfu/mL,灵敏度比直接法提高了100倍,且具有更宽的检测范围.本方法不仅检测时间短,而且具有良好的选择性和重现性.     

彩色表面等离子体共振成像初探

自主设计并建立了彩色表面等离子共振成像(Color SPRI, CSPRI)实验系统, 并结合利用自己编制的软件开展了相关研究, 成功地观测到溶液和蛋白点阵的彩色图像. 这些结果显示, CSPRI有可能成为生物分子微点阵(或生物芯片)的一种新型的彩色显示手段.     

Discovery of Novel Pleuromutilin Derivatives as Potent Antibacterial Agents for the Treatment of MRSA Infection

A series of novel pleuromutilin derivatives containing nitrogen groups on the side chain of C14 were synthesized under mild conditions. Most of the synthesized derivatives displayed potent antibacterial activities. Compound 9 was found to be the most active antibacterial derivative against MRSA (MIC = 0.06 μg/mL). Furthermore, the result of time-kill curves showed that compound 9 had a certain inhibitory effect against MRSA in vitro. Moreover, according to a surface plasmon resonance (SPR) study, compound 9 (K_D = 1.77 × 10⁻⁸ M) showed stronger affinity to the 50S ribosome than tiamulin (K_D = 2.50 × 10⁻⁸ M). The antibacterial activity of compound 9 was further evaluated in an MRSA-infected murine thigh model. Compared to the negative control group, tiamulin reduced MRSA load (~0.7 log₁₀ CFU/mL), and compound 9 performed a treatment effect (~1.3 log₁₀ CFU/mL). In addition, compound 9 was evaluated in CYP450 inhibition assay and showed only moderate in vitro CYP3A4 inhibition (IC₅₀ = 2.92 μg/mL).     

基于SPR光谱分析的液体折射率测量研究

文章从电磁场理论和射线理论的角度,详细讨论了光纤表面等离子体波传感器工作原理,分析了表面等离子体共振(SPR)光谱共振波长与液体介质折射率之间的对应关系。采用相关光谱检测技术,获得八种不同类型分析醇所对应的SPR光谱曲线族,其共振波长随液体折射率的增大而逐渐向长波长方向偏移。通过对乙醇与乙二醇混合液体所对应的共振光谱分析,实现对两者配比浓度的测定。整个传感系统结构简单,测量光路部分实现全光纤化,不仅能够对目标测量介质的实时、快速、精确测量,还可用于特殊测量场合,实现远程遥测功能。     

SPR棱镜传感器测量液体折射率的原理

推导了消失波函数的表达式,介绍了衰减全反射现象及SPR传感器测量液体折射率的原理,并利用SPR传感器测量了牛奶样品的反射系数及折射率与脂肪等主要成分含量的关系曲线.     

溶液pH对羟基苯硫酚同分异构体脱羟基反应影响的SERS研究

金属纳米结构因表面等离激元(SPR)而产生光学增强和催化效应已成为表面科学研究热点之一。SPR和电化学联用可以诱导催化一些非常规反应,并且不同pH值电解质溶液可改变表面吸附分子的存在形式,影响SPR光催化反应。以羟基苯硫酚的同分异构体为探针,采用电化学表面增强拉曼光谱(SERS)研究了取代基羟基位置、溶液pH值等对其在银电极表面吸附和SPR催化反应行为。结果表明,不同羟基取代基位置的羟基苯硫酚SPR催化脱羟基反应对溶液pH值的敏感程度不同,邻羟基苯硫酚(OHTP)的C—O键谱峰强度的变化与溶液pH值相关,其O端更易与金属作用而吸附在表面,且随pH增大而增强。对羟基苯硫酚(PHTP)在碱性条件下被完全抑制的脱羟基反应在间羟基苯硫酚(MHTP)和OHTP中均可发生。MHTP在中性(pH 7)溶液中SPR催化脱羟基反应效率最高,约为酸性(pH 2)的1.36倍,碱性(pH 12)的2.70倍。OHTP在碱性(pH 12)溶液中SPR催化脱羟基反应效率最高,约为酸性(pH 2)的13.71倍,中性(pH 7)的4.95倍。SPR催化脱羟基主要源于非去质子化条件以及形成Ag—O键这两种途径。酸性条件下MHTP及OHTP的脱羟基反应主要是未去质子化的羟基反应,碱性条件主要因去质子化后形成Ag—O键所致。中性条件下,两种贡献同时发生。对MHTP而言,由于位阻效应仅部分分子去质子化后形成Ag—O键而促进SPR催化脱羟基,因此pH 7溶液中两种效应的同时作用导致催化效率最高。对于OHTP分子,去质子化状态的O端更易与电极表面发生作用,且pH升高羟基呈现的去质子化程度更加彻底,更有利于发生脱羟基反应,在pH 12溶液中脱羟基反应主要由于形成Ag—O键,其效率亦最高。同分异构体结构以及介质酸碱度对SPR催化脱羟基反应的研究对于拓宽SPR催化反应类型及从分子水平解析其机理具有重要意义。     

基于光纤SPR光谱分析的污水降解过程监测研究

讨论了光纤表面等离子体波传感器的工作原理,并对将其用于监测以甲基橙为代表的环境污水降解过程的可行性进行了探讨。用光纤SPR传感器监测了50 mL初始浓度为30 mg·L-1的甲基橙原溶液在降解过程浓度的变化,对降解过程中光纤SPR传感器的光谱进行了详细的分析;同时采用紫外-分光光度计对降解过程中溶液浓度的变化进行了监测,并对2种方法所测的数据进行了分析对比。结果表明,光纤SPR同常规方法的测量结果一致,随着降解时间的增加,甲基橙溶液的吸光度和浓度逐渐减小,光纤SPR传感器的共振波长逐渐发生蓝移,同初始标定的甲基橙原溶液共振光谱比较,说明甲基橙逐渐被降解,且在2 h内降解率达到73%,说明用光纤SPR传感器监测污水降解过程是完全可行的。研究结果不仅为环境污水降解过程提供了一种新的监测方法,同时促进了我国SPR传感技术与环保监测研究结合,为光纤SPR技术走向实用,并最终实现产业化积累了经验。