质谱MRM技术在生物标志物研究中的应用

质谱多反应监测（multiple reaction monitoring,MRM）技术是一种基于已知信息或假定信息设定质谱检测规则,针对性的获取数据的方式,具有高选择性、高特异性、高灵敏度等优点,在基于蛋白质组学的生物标志物研究中,逐渐受到研究者的重视.从MRM定义、MRM在生物标志物研究中的优势、应用及其局限性和前景进行综述.     

糖组学技术及其在疾病标志物挖掘中的应用

蛋白质糖基化修饰的变化与癌症和其他疾病密切相关,也成为挖掘肿瘤等疾病诊疗标志物的有效手段。定量技术的精准是揭示糖组奥秘的关键前提,糖链疾病标志物的筛选和验证离不开对临床大样本的研究,因此糖组学研究和应用技术应简便、快速、高通量、准确和实用。本文对一些常用糖组分析技术特点及其在糖链标志物中的应用研究进行了简要论述,重点阐...     

人体呼出气的分析研究与临床应用进展

人体呼出气中包含的数千种挥发性有机化合物(VOC),是人体代谢和外源性化合物暴露过程的产物,外源性VOC可表征外界因素对人体健康的影响;人体代谢产生的内源性VOC是呼吸代谢组学的媒介,可作为生物标志物用于人体健康状况评估以及疾病诊断,在临床应用中具有极大的潜力;此外,很多人也开展了使用训练有素的嗅探犬进行疾病筛查的研究。在临床和实验室针对人体呼出气的研究中,主要通过无创且快速的手段进行气味样本采集,并采用基于气相色谱、质谱、激光光谱以及气体传感器等的方法进行检测分析。该文主要对人体呼出气的采集、检测分析技术、疾病生物标志物的发现以及临床应用等方面的研究进展进行综述,并对人体呼出气的分析研究与应用进行展望。     

MALDI-IMS直接组织分析

在生物医学研究中,新的生物标志物的发现和新药物的研发,需要灵敏、高通量、全面、快速的分析技术。利用基质辅助激光解吸附电离成像质谱(matrix-assisted laser desorption     

质谱技术在多组学研究和医学检验中的应用前景及挑战

传统医学模式正进入到基因组学、蛋白质组学、代谢组学等多组学整合分析的精准诊断时代。以高性能质谱为核心的多组学研究已成为各类疾病筛查、早期诊断、治疗监测和预后评估的生物标志物创新发现的关键技术平台。近年来质谱技术的迅速发展及其在临床诊断中的推广应用,为提升医学检验水平奠定了坚实的基础,临床质谱技术将是医学检验未来发展的一大亮点。本文概述了质谱及多组学在医学检验中的现状、亟待解决的瓶颈及今后的发展趋势。     

肿瘤标志物的临床应用及研究进展

自1978年Herberman在美国召开的人类免疫及肿瘤免疫诊断会上首次提出肿瘤标志物(tumor markers)概念以来,肿瘤标志物检测广泛应用于临床,在肿瘤的辅助诊断、疗效评价、复发或转移监测、预后判断方面发挥重要作用.近年来,随着生化免疫学、分子生物学、生物芯片和蛋白质组学技术的发展,肿瘤生物学标志物的临床应用取得了迅速进展,肿瘤标志物[癌基因、抑癌基因和肿瘤相关基因及其产物,循环核酸(DNA和mRNA)标志]检测及应用已进入分子和基因水平阶段.蛋白质组学成为肿瘤标志物研究的热点.血清肿瘤标志物已成为肿瘤患者实验室检查的重要指标.现就循环血液中肿瘤标志物的临床应用及研究进展作一综述.     

质谱分析在临床中的应用

质谱分析的高灵敏度、高准确度、快速、易于自动化等特点，在临床医学、基础医学和生命科学领域的应用和研究日益广泛。本文综述了近年来质谱在蛋白质、核酸、糖类、药物代谢以及微生物检验等方面的应用及进展。     

生物质谱在检验医学中的应用

1906年,英国物理学家J.J.Thomson(1906年诺贝尔物理学奖获得者)研制出了世界上第一台质谱仪器。20世纪20年代质谱才逐渐成为一种分析手段,被化学家采用。20世纪40年代,化学家认识到质谱在有机化合物结构方面能提供大量的有用信息,质谱广泛应用于有机物质分析,被喻为“一个完全的化学实验室”。直到80年代发现软电离技术[1],能用于分析高极性、难挥发和热不稳定样品气相离子化技术出现之后,生物质谱才迅速发展起来,同时推动了各种质谱联用技术的迅速发展。1质谱仪分析原理及结构简介质谱(mass spectrum,MS)是样品分子或原子在外部能量作用…     

液相色谱质谱联用技术在临床检测和代谢研究中的应用进展

色谱-质谱联用技术[液相色谱-质谱联用(LC-MS)、气相色谱-质谱联用(GC-MS)等]因其灵敏度高、具有保留时间和质荷比双重分离的选择性和特异度、检测动态范围宽等优势,已成为临床代谢物指标检测和代谢组学研究中不可或缺的分析工具。其中液相色谱串联质谱(LC-MS/MS)可对目标代谢物进行准确定性定量分析,能够在单次分析中获得数千种代谢物的信息。该文结合文献报道,综述了液质联用技术在临床样品的代谢物检测和代谢组学研究的应用进展,介绍了多种代谢标志物与疾病相关性和相应的检测方法,讨论了现阶段液质联用技术在临床转化应用中面临的问题,并展望未来发展趋势。     

呼吸道病毒实验室检测技术的研究进展

呼吸道病毒是威胁人类健康的主要因素之一,因此快速准确地鉴别病原微生物具有重要的临床意义。文章介绍了目前应用于呼吸道病毒检测的一些经典或新兴实验室技术,包括病毒分离培养技术、免疫学技术、核酸扩增技术、芯片技术、下一代测序技术、质谱(MS)技术等。这些技术以其各自的特点应用于呼吸道病毒的临床诊断、疗效观察以及科学研究与探索。     

Tandem mass spectrometry in the clinical chemistry laboratory

Tandem mass spectrometry is becoming an increasingly important analytical technology in the clinical laboratory environment. Applications in toxicology and therapeutic drug monitoring have opened the door for tandem mass spectrometry and now we are seeing a vast array of new applications being developed. It has been the combination of tandem mass spectrometry with sample introduction techniques employing atmospheric pressure ionization that has enabled this technology to be readily implemented in the clinical laboratory. Although its major research applications started with pharmacology and proteomics, tandem mass spectrometry is being used for a great variety of analyses from steroids to catecholamines to peptides. As with chromatographic methods, tandem mass spectrometry is most cost effective when groups of compounds need to be measured simultaneously. However as the price/performance of this technology continues to improve, it will become even more widely utilized for clinical laboratory applications.     

Microsatellites: perspectives and potentials of mass spectrometric analysis

Mass spectrometry is a powerful analytical tool in biotechnology. The 'soft' ionization and desorption technologies matrix-assisted laser desorption/ionization and electrospray ionization have enabled mass spectrometric analysis of large biomolecules, such as proteins and nucleic acid amplification products, and paved the way for mass spectrometry to become a leading technology in current genomics and proteomics efforts. Large-scale analysis of single nucleotide polymorphisms by mass spectrometry has been commercially established. This article reviews applications of mass spectrometry for microsatellite analysis. Features and capabilities of the two most prominent techniques, matrix assisted laser desorption/ionization and electrospray-ionization mass spectrometry, are compared and their potential to address the limitations of conventional microsatellite analysis based on comparison of gel electrophoretic mobilities is explored.     

Primer on medical genomics. Part IV: Expression proteomics

Proteomics, simply defined, is the study of proteomes. More completely, proteomics is defined as the study of all proteins, including their relative abundance, distribution, posttranslational modifications, functions, and interactions with other macromolecules, in a given cell or organism within a given environment and at a specific stage in the cell cycle. Proteins carry out the biological functions encoded by genes; hence, once the initial stage of genome sequencing and gene discovery is completed, a study of the proteome must be undertaken to address fundamental biological questions. The 3 broad areas are expression proteomics, which catalogues the relative abundance of proteins; cell-mapping or cellular proteomics, which delineates functional protein-protein interactions and organelle-specific protein distribution; and structural proteomics, which characterizes the 3-dimensional structure of proteins. With these approaches, proteins are studied on a global scale using a synergistic combination of powerful, high-throughput technologies, including 2-dimensional polyacrylamide gel electrophoresis, mass spectrometry, multidimensional liquid chromatography, and bioinformatics. Mass spectrometry, which provides highly accurate molecular mass measurements, has emerged as the analytical technology of choice for protein identification, characterization, and sequencing. This task has been made considerably easier with the availability of complete, nonredundant, and annotated genome sequence databases for many organisms. This article reviews the area of expression proteomics.     

Proteomic patterns as a diagnostic tool for early-stage cancer: a review of its progress to a clinically relevant tool.

The pace of development in novel technologies that promise improvements in the early diagnosis of disease is truly impressive. One such technology at the forefront of this revolution is mass spectrometry. New capabilities in mass spectrometry have provided the means for the development of proteomics, and the race is on to find innovative ways to apply this powerful technology to solving the problems faced in clinical medicine. One area that has garnered much attention over the past few years is the use of mass spectral patterns for cancer diagnostics. The use of these so-called 'proteomic patterns' for disease diagnosis relies fundamentally on the pattern of signals observed within a mass spectrum rather than the more conventional identification and quantitation of a biomarker such as in the case of cancer antigen-125- or prostate-specific antigen. The inherent throughput of proteomic pattern technology enables the analysis of hundreds of clinical samples per day. Currently, there are two primary means by which proteomic patterns can be acquired, surface-enhanced laser desorption/ionization (SELDI) and an electrospray ionization (ESI) method that has been popularized under the name, OvaCheck. In this review, an historical perspective on the development of proteomic patterns for the diagnosis of early-stage cancers is described. In addition, a critical assessment of the overall technology is presented with an emphasis on the steps required to enable proteomic pattern analysis to become a viable clinical tool for diagnosing early-stage cancers.     

Bioinformatics strategies for proteomic profiling

Clinical proteomics is an emerging field that involves the analysis of protein expression profiles of clinical samples for de novo discovery of disease-associated biomarkers and for gaining insight into the biology of disease processes. Mass spectrometry represents an important set of technologies for protein expression measurement. Among them, surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI TOF-MS), because of its high throughput and on-chip sample processing capability, has become a popular tool for clinical proteomics. Bioinformatics plays a critical role in the analysis of SELDI data, and therefore, it is important to understand the issues associated with the analysis of clinical proteomic data. In this review, we discuss such issues and the bioinformatics strategies used for proteomic profiling.     

Plasma proteomics-based identification of novel biomarkers in early gastric cancer

BackgroundIdentification and treatment in the early stage can significantly improve the prognosis of gastric cancer (GC). However, to date, there is still no ideal biomarker that can be used for the screening of early stage GC (EGC). The proteomics supported by mass spectrometry offers more possibilities for discovering tumor biomarkers. The aim of this study was to explore candidate protein biomarkers for EGC screening with mass spectrometry and bioinformatics technology.MethodsPlasma samples were collected from 15 EGC patients and 15 healthy controls. After a selective immune-depletion to remove high abundance proteins, plasma samples were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with the tandem mass tags (TMT) labeling.ResultsA total of 2040 proteins were identified, and 11 proteins were found to be differentially expressed. The results of the logistic regression model and orthogonal signal correction-partial least squares discriminant analysis (OPLS-DA) model showed that the changed proteins identified by plasma proteomics could help distinguish EGC patients from healthy controls.ConclusionThe proteins identified by plasma proteomics using LC-MS/MS combined with TMT labeling could help distinguish EGC from healthy controls.     

Shotgun proteomics: tools for the analysis of complex biological systems

Recent interest in proteomics has been fueled by the completion of multiple genome projects and ignited by the common need of biologists to rapidly and comprehensively evaluate complex samples of proteins on a global level. 'Shotgun proteomics' refers to the direct analysis of complex protein mixtures to rapidly generate a global profile of the protein complement within the mixture. This approach has been facilitated by the use of multidimensional protein identification technology (MudPIT), which incorporates multidimensional high-pressure liquid chromatography (LC/LC), tandem mass spectrometry (MS/MS) and database-searching algorithms. This review will focus on the most recent advances in methodologies for shotgun proteomics and address the limitations of the application of each to real biological samples.     

Enabling parallel protein analysis through mass spectrometry

The targets of the majority of drugs on the market and in development are proteins, and the efficient analysis of these molecules is critical to defining targets for better therapeutic intervention. Mass spectrometry is currently the key technology for parallel protein analysis (also referred to as proteomics). In this review we will describe recent advances in mass spectrometry instrumentation, methods and applications that are likely to impact drug discovery.     

Vascular proteomic mapping in vivo

Summary.  Molecular targeting of drugs and imaging agents remain important yet elusive goals in modern medicine. Technological advancements in genomics and proteomics methods have detected differentially expressed genes and proteins, uncovering many new candidate targets in a wide array of diseases and tissues. However, methods to validate potential targets in vivo tend to be quite laborious so that the validation and testing phase has become rate-limiting in bringing treatments to the clinic. There is a critical need for integrated approaches combining state-of-the-art methodologies in proteomics and in vivo imaging to accelerate validation of newly discovered vascular targets for nanomedicines, drugs, imaging agents, and gene vectors. This paper is a review of vascular targeting and proteomics, and will present recent developments in proteomic imaging. A new in vivo organellar proteomic imaging platform will be discussed, which combines subcellular fractionation, mass spectrometry, bioinformatic database interrogation, monoclonal antibody technology and a battery of imaging modalities to rapidly discover and validate tissue-specific endothelial protein targets in vivo . Technological advancements are permitting large-scale proteomic mapping to be performed. New targets have been discovered that permit organ-specific targeting in vivo . Improvements in imaging are creating standards for validation of targets in vivo . Tumor imaging and radioimmunotherapy have also been improved through these efforts. Although we are moving towards a comprehensive mapping of the protein expression by the endothelium, much more needs to be done.     

Serum protein fingerprinting

The core technologies in the rapidly expanding field of proteomics have matured to the point where quantitative measurements of thousands of proteins can be conducted, enabling truly global measurements of protein expression. This advent has brought with it the hope of discovering novel biomarkers that promise a renaissance in clinical medicine. To meet this need, many proteomic studies have focused on the identification and subsequent comparative analysis of the thousands of proteins that populate complex biological systems such as serum and tissues. A novel application of mass spectrometry has been in proteomic pattern analysis, which has emerged as an effective method for the early diagnosis of diseases. In stark contrast to 'classical' proteomics, proteomic pattern analysis relies on the pattern of proteins observed, rather than on the discrete identification of a protein. Proteomic pattern technology allows hundreds of clinical samples to be analyzed per day and promises to be a novel, highly sensitive predictive clinical tool to improve diagnostic and prognostic medicine.