纳米金生物探针及其应用

纳米技术与生物技术的融合,使纳米生物技术获得了很多重要的进展.纳米金作为研究较早的一种纳米材料,其在纳米生物探针方面的应用是最近几年较为活跃的研究方向.本文评述了近年来纳米金探针在生物分析中的应用及进展,阐述了纳米金与生物大分子作用的机理,介绍了纳米金探针在核酸分析、免疫分析、单细胞分析和靶向药物载体4个方面应用的最新进展,并对其未来的发展方向进行了展望.     

纳米金生物探针及其应用

纳米技术与生物技术的融合,使纳米生物技术获得了很多重要的进展.纳米金作为研究较早的一种纳米材料,其在纳米生物探针方面的应用是最近几年较为活跃的研究方向.本文评述了近年来纳米金探针在生物分析中的应用及进展,阐述了纳米金与生物大分子作用的机理,介绍了纳米金探针在核酸分析、免疫分析、单细胞分析和靶向药物载体4个方面应用的最新进展,并对其未来的发展方向进行了展望.     

肿瘤病理可视化纳米探针的研究进展※

癌症的发生和发展伴随着一系列复杂的分子病理学变化, 具有极大的个体差异性. 因此, 实现肿瘤的精准诊断, 尤其是分子病理学的诊断尤为重要. 在临床检测中, 传统影像学检查可以反映肿瘤的位置和解剖学结构, 却难以对其分子病理做出判定; 而病理活检虽然可以获取肿瘤的分子学特征, 但需通过创伤性手段获取样本, 且具有时空局限性. 相比之下, 借助于特异性探针成像的肿瘤分子影像学, 直接以肿瘤病理分子标志物作为成像对比度的来源, 旨在从分子层面对肿瘤进展中的病理学特征进行在体定量化分析, 在肿瘤的精准诊断中具备独特的优势. 近年来, 纳米材料由于优越的理化性质, 已经成为构建高灵敏肿瘤分子影像探针的重要信号载体之一. 基于此, 本综述从基本的纳米靶向探针, 到光、磁学智能响应型纳米探针, 系统总结归纳了基于纳米材料的分子影像技术对肿瘤病理在体可视化的研究进展, 并对未来临床环境中实施该纳米探针技术进行了展望.     

纳米探针芯片技术用于微量乙肝病毒DNA的检测

利用两组探针修饰的微粒:(1)表面标记有可与待测乙肝病毒(HBV) DNA另一端结合的纳米金探针1(信号探针)以及可与信号探针部分结合的纳米金探针2(检测探针);(2)表面标记有可与待测HBV DNA一端结合的磁珠探针(捕捉探针1).检测靶HBV DNA时,磁珠探针与信号探针在液相中可分别与HBV DNA靶序列一端结合最终形成三明治样结构.再以磁场将三明治样复合物从反应液中分离,以DTT溶液将信号探针从纳米金颗粒上洗脱.洗脱后的信号探针数量反映靶基因的多寡,信号探针一段与预先点样的基因芯片上的捕捉探针2结合,检测探针与信号探针另一段相结合,最后用银染液将检测探针显色从而得到靶目标DNA相对定量信息.结果表明,本检测方法的检测灵敏度达到10-15 mol/L水平.检测时间少于1.5 h,检测结果与HBV DNA水平呈现较好的线性关系且无假阳性结果;本方法有望用于乙肝病人血清中HBV DNA的快速筛测及其它微生物基因的检测.     

自循环稀土纳米荧光探针用于生物样本中抗坏血酸检测

通过化学键连法在氨基化SiO₂包覆的NaLuF₄:Yb,Er@NaLuF₄核壳结构纳米探针表面修饰水杨酸-铁配合物制备得到自循环稀土纳米荧光探针。观察到探针在540和1530 nm两处荧光发射峰的比值随抗坏血酸浓度增高而增大,并在0～1.0 mmol·L^-1浓度范围内呈现良好的线性关系。将与抗坏血酸反应后的探针静置一定时间后离心分离,可实现探针的循环利用,并在10次循环内保证荧光信号比值恢复至与抗坏血酸反应前的水平(比值变化<5%)。通过构建小鼠模型,并使用这一探针测定其血清和尿液中抗坏血酸浓度,初步验证了评估生物样本中抗坏血酸水平的可行性。     

基于生物功能化纳米DNA探针及其传感策略

随着人类基因组计划的完成和功能基因研究的深入,基因诊断已成为分子生物学和生物医学的重要研究领域。DNA生物传感器作为一种利用核酸碱基配对原则进行识别,能对基因片段实现持续、快速、灵敏和选择性检测的新方法,近年来发展非常迅速。纳米材料由于具有独特物理化学性质、良好的生物相容性、优越的机械性能及表面易于生物功能化等特点,被广泛应用到生物分析之中。各种各样组成、尺寸、维度及形状的纳米材料如量子点、贵金属纳米材料、碳纳米材料等被可控地修饰上不同的生物分子,用于发展特殊性质的纳米探针,构建DNA生物传感器,实现对DNA片段高灵敏及高特异性的检测。     

DNA功能化纳米金用于生物小分子检测

纳米金是金的微小颗粒,在水溶液中以胶体金的形态存在。胶体金的颜色会随着其粒径及表面修饰差异而发生变化,这种颜色变化可通过肉眼观察;同时,这种改变会产生强烈的光散射或光吸收信号。基于这种信号而建立的纳米金比色检测法,已被广泛用于生物分子(如核酸、蛋白质、多糖甚至是细胞)的检测。DNA功能化纳米生物传感器是利用核酸碱基配对原则进行识别,能实现特定基因片段的持续、快速、灵敏和选择性检测。本文结合最近十年的研究现状,主要论述了DNA功能化纳米金用于比色检测法的原理及用于核酸、蛋白质和部分生物小分子的检测,并评述了其中的挑战和前景。     

DNA功能化纳米金用于生物小分子检测

纳米金是金的微小颗粒,在水溶液中以胶体金的形态存在。胶体金的颜色会随着其粒径及表面修饰差异而发生变化,这种颜色变化可通过肉眼观察;同时,这种改变会产生强烈的光散射或光吸收信号。基于这种信号而建立的纳米金比色检测法,已被广泛用于生物分子(如核酸、蛋白质、多糖甚至是细胞)的检测。DNA功能化纳米生物传感器是利用核酸碱基配对原则进行识别,能实现特定基因片段的持续、快速、灵敏和选择性检测。本文结合最近十年的研究现状,主要论述了DNA功能化纳米金用于比色检测法的原理及用于核酸、蛋白质和部分生物小分子的检测,并评述了其中的挑战和前景。     

研究发现纳米金刚石可杀菌

<正>德国不来梅大学研究人员参与的一个国际研究团队发现,纳米金刚石可像金属银、铜一样有效杀除细菌。纳米金刚石直径约5纳米(1纳米等于10亿分之1米),约为细菌的二百分之一,可通过含碳化合物在高压容器中爆炸产生。这种灰褐色金刚石粉末在接受不同的热处理后,表面会形成不同的化学基团。研究人员发现,一些纳米金刚石具有较强的杀菌特性,可在短时间内杀死细菌,而纳米金刚石表面上一种名为酸酐的特定含氧基团似乎是其具有杀菌特性的原因所在。     

Recent Applications of Aggregation Induced Emission Probes for Antimicrobial Peptide Studies

Antimicrobial peptides (AMPs) are being intensively investigated as they are considered promising alternatives to antibiotics where their clinical efficacy is dwindling due to the emergence of antimicrobial resistance (AMR). Accompanying with the development of AMPs, a number of fluorescent probes have been developed to facilitate the understanding the modes of action of AMPs. These probes have been used to monitor the binding process, determine the working mechanism and evaluate the antimicrobial properties of AMPs. In particular, with the recent advance of aggregation-induced emission (AIE) fluorophores, that show many advantageous properties over traditional probes, there is an increasing research interest in using AIE probes for AMP studies. In this review, we give an overview of AMP development, highlight the recent progress of using fluorescence probes in particularly AIE probes in the AMP field and propose the future perspective of developing potent antimicrobial agents to combat AMR.     

Fluorescent Self-Threaded Peptide Probes for Biological Imaging

A general synthetic method creates a new class of covalently connected, self-threaded, fluorescent molecular probes with figure-eight topology, an encapsulated deep-red fluorophore, and two peripheral peptide loops. The globular molecular shape and rigidified peptide loops enhance imaging performance by promoting water solubility, eliminating probe self-aggregation, and increasing probe stability. Moreover, the peptide loops determine the affinity and selectivity for targets within complex biological samples such as cell culture, tissue histology slices, or living subjects. For example, a probe with cell-penetrating peptide loops targets the surface of cell plasma membranes, whereas, a probe with bone-targeting peptide loops selectively stains the skeleton within a living mouse. The unique combination of bright deep-red fluorescence, high stability, and predictable peptide-based targeting is ideal for photon intense fluorescence microscopy and biological imaging.     

Polymeric 1H MRI Probes for Visualizing Tumor In Vivo

Magnetic resonance imaging (MRI) has become a prominent non– or low–invasive imaging technique, providing high–resolution, three–dimensional images as well as physiological information about tissues. Low–molecular–weight Gd–MRI contrast agents (CAs), such as Gd–DTPA (DTPA: diethylenetriaminepentaacetic acid), are commonly used in the clinical diagnosis, while macromolecular Gd–MRI CAs have several advantages over low–molecular–weight Gd–MRI CAs, which help minimize the dose of CAs and the risk of side effects. Accordingly, we developed chiral dendrimer Gd–MRI CAs, which showed high r₁ values. The association constant values (K_a) of S–isomeric dendrimer CAs to bovine serum albumin (BSA) were higher than those of R–isomeric dendrimer CAs. Besides, based on a totally new concept, we developed ¹³C/¹⁵N–enriched multiple–resonance NMR/MRI probes, which realized highly selective observation of the probes and analysis of metabolic reactions of interest. This account summarizes our recent study on developing both chiral dendrimer Gd–MRI CAs, and self–traceable ¹³C/¹⁵N–enriched phosphorylcholine polymer probes for early detection of tumors.     

Multiple Mechanisms of the Synthesized Antimicrobial Peptide TS against Gram-Negative Bacteria for High Efficacy Antibacterial Action In Vivo

The emergence of drug-resistant bacteria emphasizes the urgent need for novel antibiotics. The antimicrobial peptide TS shows extensive antibacterial activity in vitro and in vivo, especially in gram-negative bacteria; however, its antibacterial mechanism is unclear. Here, we find that TS without hemolytic activity disrupts the integrity of the outer bacterial cell membrane by displacing divalent cations and competitively binding lipopolysaccharides. In addition, the antimicrobial peptide TS can inhibit and kill E. coli by disintegrating the bacteria from within by interacting with bacterial DNA. Thus, antimicrobial peptide TS’s multiple antibacterial mechanisms may not easily induce bacterial resistance, suggesting use as an antibacterial drug to be for combating bacterial infections in the future.     

液态石油烃的脱砷及砷化物与亚铜探针的相互作用

采用密度泛函理论B3LYP方法, 在6-311+G(2df,2p)基组水平上研究了液态石油烃体系中甲基胂化合物与过渡金属探针Cu+和CuCl的相互作用. 结果表明, 甲基胂化物与亚铜离子作用的最稳定模式为四面体构型, 随着烃基取代数增加, 砷化物同Cu+或CuCl的相互作用能(E0)更负, 配合物更稳定, 同时二者的相互作用轨道的能级差(△E)与E0值线性相关(R2≥0.99). Cu+配合物中, As—Cu成键轨道向C—As和/或H—As反键轨道形成电子反馈, 但是CuCl配合物中类似电子反馈则没有形成. 烃基取代并不降低活性组分对砷化物的吸附活性, 在活性组分满足砷化物选择性吸附的条件下, 液态石油烃脱砷的主要控制因素为砷化物在脱砷剂内部孔道中的扩散传输. 有机硫化物噻吩并不影响砷化物的选择性吸附分离, 硫醇类化合物则会影响单烷基砷化物的选择性吸附. 在进行脱砷剂开发时, 应拓宽载体孔道以提高流体传输效率, 并选择活性相组分与砷化物作用的△E相对较小, 配合物容易形成电子反馈, 且对硫醇类化合物的作用能力相对较弱, 对单烷基砷化物的作用能力相对较强的过渡金属组分.     

Recent Advances in the Application of the Antimicrobial Peptide Nisin in the Inactivation of Spore-Forming Bacteria in Foods

Conventional thermal and chemical treatments used in food preservation have come under scrutiny by consumers who demand minimally processed foods free from chemical agents but microbiologically safe. As a result, antimicrobial peptides (AMPs) such as bacteriocins and nisin that are ribosomally synthesised by bacteria, more prominently by the lactic acid bacteria (LAB) have appeared as a potent alternative due to their multiple biological activities. They represent a powerful strategy to prevent the development of spore-forming microorganisms in foods. Unlike thermal methods, they are natural without an adverse impact on food organoleptic and nutritional attributes. AMPs such as nisin and bacteriocins are generally effective in eliminating the vegetative forms of spore-forming bacteria compared to the more resilient spore forms. However, in combination with other non-thermal treatments, such as high pressure, supercritical carbon dioxide, electric pulses, a synergistic effect with AMPs such as nisin exists and has been proven to be effective in the inactivation of microbial spores through the disruption of the spore structure and prevention of spore outgrowth. The control of microbial spores in foods is essential in maintaining food safety and extension of shelf-life. Thus, exploration of the mechanisms of action of AMPs such as nisin is critical for their design and effective application in the food industry. This review harmonises information on the mechanisms of bacteria inactivation from published literature and the utilisation of AMPs in the control of microbial spores in food. It highlights future perspectives in research and application in food processing.     

Acidic‐pH‐Activatable Fluorescence Probes for Visualizing Exocytosis Dynamics

Live imaging of exocytosis dynamics is crucial for a precise spatiotemporal understanding of secretion phenomena, but current approaches have serious limitations. We designed and synthesized small‐molecular fluorescent probes that were chemically optimized for sensing acidic intravesicular pH values, and established that they can be used to sensitively and reliably visualize vesicular dynamics following stimulation. This straightforward technique for the visualization of exocytosis as well as endocytosis/reacidification processes with high spatiotemporal precision is expected to be a powerful tool for investigating dynamic cellular phenomena involving changes in the pH value.     

谷胱甘肽和凋亡酶-3响应的环肽分子荧光探针

设计、合成了一类新型谷胱甘肽(glutathione,GSH)和凋亡酶-3(Caspase-3)响应的环肽分子荧光探针.该类探针主要由能量共振转移(FRET)分子荧光对、Caspase-3特异性识别多肽序列和GSH响应双硫键组成,分为不含穿膜肽序列(CP)和包含穿膜肽序列(cp CP)的两种不同环肽分子荧光探针.2种环肽分子荧光探针均能实现在GSH和Caspase-3同时存在情况下的精确成像,同时具有良好的响应性、特异性和高信噪比.该类环肽分子荧光探针在细胞培养环境中具有良好的稳定性和生物相容性.利用该探针,可以实现对星形孢菌素(STS)诱发的细胞凋亡进行实时、原位的成像监测,并对抗肿瘤药物阿霉素(DOX)和顺铂(cisplatin)诱导的细胞凋亡进行成像.这种具有多重响应并能用于精确成像的分子荧光探针将极大地促进疾病的精确诊断.