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25 Years Responding to Respiratory and Other Viruses with Mass Spectrometry 25年来对呼吸道和其他病毒的质谱反应
Q3 Physics and Astronomy Pub Date : 2023-01-01 DOI: 10.5702/massspectrometry.a0136
Kevin M. Downard
This review article presents the development and application of mass spectrometry (MS) approaches, developed in the author’s laboratory over the past 25 years, to detect; characterise, type and subtype; and distinguish major variants and subvariants of respiratory viruses such as influenza and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). All features make use of matrix-assisted laser desorption ionisation (MALDI) mass maps, recorded for individual viral proteins or whole virus digests. A MALDI-based immunoassay in which antibody–peptide complexes were preserved on conventional MALDI targets without their immobilisation led to an approach that enabled their indirect detection. The site of binding, and thus the molecular antigenicity of viruses, could be determined. The same approach was employed to study antivirals bound to their target viral protein, the nature of the binding residues, and relative binding affinities. The benefits of high-resolution MS were exploited to detect sequence-conserved signature peptides of unique mass within whole virus and single protein digests. These enabled viruses to be typed, subtyped, their lineage determined, and variants and subvariants to be distinguished. Their detection using selected ion monitoring improved analytical sensitivity limits to aid the identification of viruses in clinical specimens. The same high-resolution mass map data, for a wide range of viral strains, were input into a purpose-built algorithm (MassTree) in order to both chart and interrogate viral evolution. Without the need for gene or protein sequences, or any sequence alignment, this phylonumerics approach also determines and displays single-point mutations associated with viral protein evolution in a single-tree building step.
本文综述了作者实验室在过去25年中开发的质谱(MS)方法的发展和应用,用于检测;特征、类型和亚型;区分流感、SARS-CoV-2等呼吸道病毒的主要变异和亚变异。所有特征都利用基质辅助激光解吸电离(MALDI)质量图,记录单个病毒蛋白或整个病毒消化。一种基于MALDI的免疫分析方法,将抗体-肽复合物保存在传统MALDI靶标上,而不固定它们,从而使它们能够间接检测。结合位点,从而确定病毒的分子抗原性。同样的方法被用于研究抗病毒药物与其靶病毒蛋白的结合、结合残基的性质和相对结合亲和力。高分辨率质谱的优势被用于检测整个病毒和单个蛋白质消化的独特质量的序列保守的特征肽。这使得病毒能够被分型、分亚型、确定它们的谱系、区分变异和亚变异。它们的检测使用选定的离子监测提高了分析灵敏度限制,以帮助鉴定临床标本中的病毒。相同的高分辨率海量地图数据,用于广泛的病毒株,被输入到一个专门构建的算法(masstreet)中,以便绘制和询问病毒的进化。不需要基因或蛋白质序列,也不需要任何序列比对,这种系统数量学方法还可以在单树构建步骤中确定和显示与病毒蛋白质进化相关的单点突变。
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引用次数: 0
Imaging Analysis of Phosphatidylcholines and Diacylglycerols Using Surface-Assisted Laser Desorption/Ionization Mass Spectrometry with Metal Film Formed by Mist Chemical Vapor Deposition 用表面辅助激光解吸/电离质谱法对磷脂酰胆碱和二酰基甘油的成像分析
Q3 Physics and Astronomy Pub Date : 2023-01-01 DOI: 10.5702/massspectrometry.a0135
Riko Takata, Yuji Nakabayashi, Kotaro Hashimoto, Akio Miyazato, Issey Osaka
Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) has been widely used for analyses of biomolecules and industrial materials. Surface-assisted laser desorption/ionization (SALDI) is studied to complement the ionization ability for the MALDI/MS. In this study, lab-made mist chemical vapor deposition (mist CVD) system was used to produce metal films as ionization assistance materials for SALDI/MS. The system could give Ag film from inexpensive silver trifluoroacetate solution rapidly and simply under atmospheric pressure. Phosphatidylcholines could be detected high sensitively and diacylglycerols (DAGs) could not be detected in MALDI/MS. In the SALDI/MS and the MS imaging with Ag film by mist CVD, both the phosphatidylcholines and the DAGs could be detected and the localized images. In the Ag film-SALDI/MS of lipids, not only Ag-adducted ions but also Na- and K-adducted ions were detected. The Ag film formed by the mist CVD to act as an ionization-assistance material and a cationization agent in SALDI would be useful in MS imaging of biological tissue sections.
基质辅助激光解吸/电离(MALDI)质谱法(MS)已广泛应用于生物分子和工业材料的分析。研究了表面辅助激光解吸/电离(SALDI),以补充MALDI/MS的电离能力。本研究采用实验室自制的雾状化学气相沉积(mist CVD)系统制备金属薄膜作为SALDI/MS的电离辅助材料。该系统可以在常压下快速、简单地从廉价的三氟乙酸银溶液中制备银膜。MALDI/MS对磷脂酰胆碱的检测灵敏度较高,对二酰基甘油的检测灵敏度较低。在SALDI/MS和雾CVD银膜MS成像中,均能检测到磷脂酰胆碱和dag,并能定位图像。在脂类的Ag膜- saldi /MS中,不仅检测到Ag-内合离子,还检测到Na-和k -内合离子。薄雾CVD形成的Ag膜在SALDI中作为电离辅助材料和阳离子化剂,可用于生物组织切片的MS成像。
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引用次数: 0
Evaluation for Ion Heating of H2A-H2B Dimer in Ion Mobility Spectrometry-Mass Spectrometry. 离子迁移率质谱法中H2A-H2B二聚体离子加热的评价。
Q3 Physics and Astronomy Pub Date : 2023-01-01 Epub Date: 2023-10-17 DOI: 10.5702/massspectrometry.A0131
Kazumi Saikusa, Daiki Asakawa, Sotaro Fuchigami, Satoko Akashi

Ion mobility spectrometry-mass spectrometry (IMS-MS) provides m/z values and collision cross sections (CCSs) of gas-phase ions. In our previous study, an intrinsically disordered protein, the H2A-H2B dimer, was analyzed using IMS-MS, resulting in two conformational populations of CCS. Based on experimental and theoretical approaches, this resulted from a structural diversity of intrinsically disordered regions. We predicted that this phenomenon is related to ion heating in the IMS-MS instrument. In this study, to reveal the effect of ion heating from parameters in the IMS-MS instrument on the conformational population of the H2A-H2B dimer, we investigated the arrival time distributions of the H2A-H2B dimer by changing values of three instrumental parameters, namely, cone voltage located in the first vacuum chamber, trap collision energy (trap CE) for tandem mass spectrometry, and trap bias voltage for the entrance of IMS. These results revealed that the two populations observed for the H2A-H2B dimer were due to the trap bias voltage. Furthermore, to evaluate the internal energies of the analyte ions with respect to each parameter, benzylpyridinium derivatives were used as temperature-sensitive probes. The results showed that the trap CE voltage imparts greater internal energy to the ions than the trap bias voltage. In addition, this slight change in the internal energy caused by the trap bias voltage resulted in the structural diversity of the H2A-H2B dimer. Therefore, the trap bias voltage should be set with attention to the properties of the analytes, even if the effect of the trap bias voltage on the internal energy is negligible.

离子迁移率谱-质谱(IMS-MS)提供气相离子的m/z值和碰撞截面(CCS)。在我们之前的研究中,使用IMS-MS分析了一种本质紊乱的蛋白质H2A-H2B二聚体,从而产生了CCS的两个构象群体。基于实验和理论方法,这是由于内在无序区域的结构多样性造成的。我们预测这种现象与IMS-MS仪器中的离子加热有关。在本研究中,为了从IMS-MS仪器中的参数揭示离子加热对H2A-H2B二聚体构象群体的影响,我们通过改变三个仪器参数的值,即位于第一真空室的锥电压、用于串联质谱的陷阱碰撞能量(陷阱CE),以及用于IMS入口的陷阱偏置电压。这些结果表明,对于H2A-H2B二聚体观察到的两个群体是由于陷阱偏置电压。此外,为了评估分析物离子相对于每个参数的内能,使用苄基吡啶鎓衍生物作为温度敏感探针。结果表明,陷阱CE电压比陷阱偏置电压赋予离子更大的内能。此外,由陷阱偏压引起的内能的这种轻微变化导致H2A-H2B二聚体的结构多样性。因此,即使陷阱偏置电压对内能的影响可以忽略不计,也应注意分析物的性质来设置陷阱偏置电压。
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引用次数: 0
Carotenoids: Distribution, Function in Nature, and Analysis Using LC-Photodiode Array Detector (DAD)-MS and MS/MS System. 类胡萝卜素:分布、在自然界中的作用和使用LC光电二极管阵列检测器(DAD)-MS和MS/MS系统的分析。
Q3 Physics and Astronomy Pub Date : 2023-01-01 Epub Date: 2023-11-01 DOI: 10.5702/massspectrometry.A0133
Takashi Maoka

Carotenoids are tetraterpene pigments that are present in photosynthetic bacteria, some species of archaea and fungi, algae, plants, and animals. Carotenoids are essential pigments in photosynthetic organs along with chlorophylls. Carotenoids also act as photo-protectors, antioxidants, color attractants, and precursors of plant hormones in plants. Carotenoids in animals play important roles, such as precursors of vitamin A, photo-protectors, antioxidants, enhancers of immunity, and contributors to reproduction. More than 850 kinds of carotenoids are present in nature. The structures are similar and all of them are labile. Analysis of natural carotenoids requires the establishment of reliable methods for analyzing them. Liquid chromatography-mass spectrometry (LC-MS) and mass spectrometry/mass spectrometry (MS/MS) coupled with photodiode array detector (DAD) is an important tool for analysis of natural carotenoids. Electrospray ionization and atmospheric pressure chemical ionization are commonly used for ionization of LC-MS of carotenoids. MS and MS/MS provide not only molecular weight information but also some structural information on carotenoids. Ultraviolet-visible spectra from DAD provide information on chromophore systems, which cannot be provided by MS spectral data. In the present review, I report the structural diversity and function of natural carotenoids, and also describe the techniques for analysis of natural carotenoids using the LC-DAD-MS and MS/MS system.

类胡萝卜素是四萜类色素,存在于光合细菌、某些古菌和真菌、藻类、植物和动物中。类胡萝卜素与叶绿素一起是光合器官中的必需色素。类胡萝卜素还起到光保护剂、抗氧化剂、颜色引诱剂和植物激素前体的作用。动物体内的类胡萝卜素发挥着重要作用,如维生素A的前体、光保护剂、抗氧化剂、免疫增强剂和生殖因子。自然界中存在850多种类胡萝卜素。这些结构是相似的,并且都是不稳定的。天然类胡萝卜素的分析需要建立可靠的分析方法。液相色谱-质谱法(LC-MS)和质谱/质谱联用光电二极管阵列检测器(DAD)是分析天然类胡萝卜素的重要工具。电喷雾电离和大气压化学电离常用于类胡萝卜素的LC-MS电离。MS和MS/MS不仅提供了类胡萝卜素的分子量信息,而且还提供了一些结构信息。DAD的紫外-可见光谱提供了发色团系统的信息,而MS光谱数据无法提供这些信息。在本综述中,我报道了天然类胡萝卜素的结构多样性和功能,并描述了使用LC-DAD-MS和MS/MS系统分析天然类胡萝卜蛋白的技术。
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引用次数: 0
Comments on "Identification of Negative Ion at m/z 20 Produced by Atmospheric Pressure Corona Discharge Ionization under Ambient Air". 关于 "常压电晕放电电离在环境空气中产生的 m/z 20 负离子的鉴定 "的评论
Q3 Physics and Astronomy Pub Date : 2023-01-01 Epub Date: 2023-12-28 DOI: 10.5702/massspectrometry.A0140
Daiki Asakawa, Kenzo Hiraoka
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引用次数: 0
Recent Advances and Applications of Ambient Mass Spectrometry Imaging in Cancer Research: An Overview. 环境质谱成像在癌症研究中的最新进展和应用综述。
Q3 Physics and Astronomy Pub Date : 2023-01-01 Epub Date: 2023-09-28 DOI: 10.5702/massspectrometry.A0129
Bharath S Kumar

Cancer metabolic variability has a significant impact on both diagnosis and treatment outcomes. The discovery of novel biological indicators and metabolic dysregulation, can significantly rely on comprehension of the modified metabolism in cancer, is a research focus. Tissue histology is a critical feature in the diagnostic testing of many ailments, such as cancer. To assess the surgical margin of the tumour on patients, frozen section histology is a tedious, laborious, and typically arbitrary method. Concurrent monitoring of ion images in tissues facilitated by the latest advancements in mass spectrometry imaging (MSI) is far more efficient than optical tissue image analysis utilized in conventional histopathology examination. This article focuses on the "desorption electrospray ionization (DESI)-MSI" technique's most recent advancements and uses in cancer research. DESI-MSI can provide wealthy information based on the variances in metabolites and lipids in normal and cancerous tissues by acquiring ion images of the lipid and metabolite variances on biopsy samples. As opposed to a systematic review, this article offers a synopsis of the most widely employed cutting-edge DESI-MSI techniques in cancer research.

癌症代谢变异性对诊断和治疗结果都有显著影响。新的生物学指标和代谢失调的发现,在很大程度上依赖于对癌症代谢改变的理解,是研究的重点。组织组织学是许多疾病(如癌症)诊断测试的关键特征。为了评估患者的肿瘤手术边缘,冷冻切片组织学是一种乏味、费力且典型的任意方法。质谱成像(MSI)的最新进展促进了对组织中离子图像的同时监测,其效率远高于传统组织病理学检查中使用的光学组织图像分析。本文重点介绍了“解吸电喷雾电离(DESI)-MSI”技术在癌症研究中的最新进展和应用。DESI-MSI可以通过获取活检样本上脂质和代谢产物变化的离子图像,提供基于正常和癌组织中代谢产物和脂质变化的丰富信息。与系统综述相反,本文简要介绍了癌症研究中最广泛使用的尖端DESI-MSI技术。
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引用次数: 0
Estimation of the Spatial Extent of the Transient Gain Drop in a Microchannel Plate. 微通道板瞬态增益下降空间范围的估计。
Q3 Physics and Astronomy Pub Date : 2023-01-01 Epub Date: 2023-11-07 DOI: 10.5702/massspectrometry.A0134
Hiroshi Kobayashi, Toshinobu Hondo, Yasuo Kanematsu, Motohiro Suyama, Michisato Toyoda

The gain of the microchannel plate temporally drops after an ion initiates an electron avalanche. Electron multiplication was expected to deplete the charge from the microchannel wall and produce the depleted charge (wall charge). Moreover, it was reported that the gain drop occurred not only in the activated channels, where the electrons are multiplied, but also in the surrounding channels. One mechanism of the gain-drop spatial extension has been considered as that the wall charges in the activated channels change the electric field in the surrounding channels. Anacker et al. assumed that the wall charge is a uniform line charge; the gain-drop spatial extent should be proportional to the amount of the wall charges. We considered that the wall charges exponentially increased in the channel toward the exit. In this study, the electric field produced by the wall charges was calculated, considering the distribution of the wall charges. The transverse electric field generated by the wall charges was expected to disturb the electron trajectory near the channel exit and decrease the number of secondary electrons emitted per collision (gain per collision), resulting in a gain drop. The gain per collision was calculated to decrease by 22% for the position where the gain decreased significantly in the presence of the transverse electric field of 3×105 V/m. In our model, the gain-drop spatial extent extended proportionally to the square root of the wall charges when the distance from the activated channel exceeded 50 μm.

离子引发电子雪崩后,微通道板的增益暂时下降。预计电子倍增会耗尽来自微通道壁的电荷并产生耗尽电荷(壁电荷)。此外,据报道,增益下降不仅发生在电子倍增的激活通道中,而且发生在周围的通道中。增益-下降空间扩展的一种机制被认为是激活通道中的壁电荷改变了周围通道中的电场。Anacker等人假设壁面电荷为均匀线电荷;增益-下降空间范围应与壁面电荷量成正比。我们认为壁面电荷沿通道向出口呈指数增长。在本研究中,考虑壁面电荷的分布,计算壁面电荷产生的电场。壁电荷产生的横向电场预计会干扰通道出口附近的电子轨迹,减少每次碰撞发射的二次电子数量(每次碰撞增益),导致增益下降。当横向电场强度为3×105 V/m时,每次碰撞的增益降低22%。在我们的模型中,当与激活通道的距离超过50 μm时,增益下降的空间范围与壁面电荷的平方根成正比。
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引用次数: 0
Fragmentation and Ionization Efficiency of Positional and Functional Isomers of Paeoniflorin Derivatives in Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry 基质辅助激光解吸/电离飞行时间质谱分析芍药苷衍生物位置和功能异构体的碎片化和电离效率
Q3 Physics and Astronomy Pub Date : 2022-02-28 DOI: 10.5702/massspectrometry.A0101
T. Yamagaki, Kohtaro Sugahara, K. Fujikawa, Kazuto Washida
Paeoniflorin and albiflorin, which are functional isomers, are the major constituents of an herbal medicine derived from Paeonia lactiflora. Those functional isomers and their galloylated derivatives, which are positional isomers, were studied by matrix-assisted laser desorption/ionization–tandem mass spectrometry (MALDI-MS/MS). The resulting mass spectra are discussed based on the fragmentation patterns of the sodium adducts. The product ion spectra of 4-O-galloylalbiflorin and 4′-O-galloylpaeoniflorin differed, even though they were positional isomers. The fragmentations of the ester parts were influenced by the neighboring hydroxyl groups. The ionization efficiency of the sodium adduct of albiflorin was higher than that for paeoniflorin. These results indicate that the carboxylic ester group has a higher affinity for sodium ions than the acetal group, which can be attributed to the carbonyl oxygen being negatively polarized, allowing it to function as a Lewis base.
芍药苷和芍药苷是芍药中功能异构体的主要成分。采用基质辅助激光解吸/电离-串联质谱(MALDI-MS/MS)对这些功能异构体及其没食子酸衍生物进行了研究。根据钠加合物的破碎模式对质谱进行了讨论。4- o -没食子碱芍药苷和4 ' - o -没食子碱芍药苷虽然是位置异构体,但它们的产物离子光谱存在差异。酯部分的断裂受邻近羟基的影响。芍药苷钠加合物的电离效率高于芍药苷。这些结果表明,羧酸酯基团比缩醛基团对钠离子具有更高的亲和力,这可能是由于羰基氧被负极化,使其具有路易斯碱的功能。
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引用次数: 0
Mass Spectrometry Imaging 质谱成像
Q3 Physics and Astronomy Pub Date : 2022-02-25 DOI: 10.5702/massspectrometry.A0102
S. Shimma
Mass spectrometry imaging (MSI) is a technique for obtaining information on the distribution of various molecules by performing mass spectrometry directly on the sample surface. The applications range from small molecules such as lipids to large molecules such as proteins. It is also possible to detect pharmaceuticals and elemental isotopes in interstellar matter. This review will introduce various applications of MSI with examples.
质谱成像(MSI)是一种通过直接在样品表面进行质谱分析来获取各种分子分布信息的技术。应用范围从小分子如脂类到大分子如蛋白质。在星际物质中探测药物和元素同位素也是可能的。本文将通过实例介绍MSI的各种应用。
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引用次数: 1
Electrospray Ionization Mass Spectrometry of Apolipoprotein CIII to Evaluate O-glycan Site Occupancy and Sialylation in Congenital Disorders of Glycosylation. 利用载脂蛋白 CIII 的电喷雾离子化质谱法评估先天性糖基化紊乱的 O-聚糖位点占用率和 Sialylation。
Q3 Physics and Astronomy Pub Date : 2022-01-01 Epub Date: 2022-08-10 DOI: 10.5702/massspectrometry.A0104
Yoshinao Wada, Nobuhiko Okamoto

Congenital disorders of glycosylation (CDG) are inherited metabolic diseases that affect the synthesis of glycoconjugates. Defects in mucin-type O-glycosylation occur independently or in combination with N-glycosylation disorders, and the profiling of the O-glycans of apolipoprotein CIII (apoCIII) by mass spectrometry (MS) can be used to support a diagnosis. The biomarkers are site occupancy and sialylation levels, which are indicated by the content of non-glycosylated apoCIII0a isoform and by the ratio of monosialylated apoCIII1 to disialylated apoCIII2 isoforms, respectively. In this report, electrospray ionization (ESI) quadrupole MS of apoCIII was used to identify these biomarkers. Among the instrumental parameters, the declustering potential (DP) induced the fragmentation of the O-glycan moiety including the Thr-GalNAc linkage, resulting in an increase in apoCIII0a ions. This incurs the risk of creating a false positive for reduced site occupancy. The apoCIII1/apoCIII2 ratio was substantially unchanged despite some dissociation of sialic acids. Therefore, appropriate DP settings are especially important when transferrin, which requires a higher DP, for N-glycosylation disorders is analyzed simultaneously with apoCIII in a single ESI MS measurement. Finally, a reference range of diagnostic biomarkers and mass spectra of apoCIII obtained from patients with SLC35A1-, TRAPPC11-, and ATP6V0A2-CDG are presented.

先天性糖基化紊乱(CDG)是一种遗传性代谢疾病,会影响糖苷酸的合成。粘蛋白型 O-糖基化缺陷可单独发生,也可与 N-糖基化紊乱同时发生,通过质谱(MS)分析载脂蛋白 CIII(apolipoprotein CIII)的 O-糖可用于辅助诊断。生物标志物是位点占有率和糖基化水平,分别通过非糖基化载脂蛋白 CIII0a 同工型的含量和单糖基化载脂蛋白 CIII1 与双糖基化载脂蛋白 CIII2 同工型的比例来表示。本报告采用电喷雾离子化(ESI)四极杆质谱对apoCIII进行了鉴定。在仪器参数中,解聚电位(DP)诱导了包括 Thr-GalNAc 连接在内的 O-糖分子的碎裂,导致 apoCIII0a 离子增加。这有可能造成位点占有率降低的假阳性结果。尽管有一些硅酸解离,但apoCIII1/apoCIII2 的比率基本未变。因此,在一次 ESI MS 测量中同时分析因 N-糖基化紊乱而需要较高 DP 值的转铁蛋白和 apoCIII 时,适当的 DP 值设置尤为重要。最后,介绍了从 SLC35A1-、TRAPPC11- 和 ATP6V0A2-CDG 患者身上获得的诊断生物标记物的参考范围和 apoCIII 的质谱。
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引用次数: 0
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Mass spectrometry
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