基于质谱的蛋白质组学技术在神经退行性疾病生物标志物研究中的应用

蛋白质组学是在整体水平上研究细胞、组织或生物体蛋白质组成及变化规律的科学.与传统的生物学研究相比,蛋白质组学具有快速、灵敏、高通量的优点.神经退行性疾病是一类由神经系统内特定神经细胞的进程性病变或丢失而导致神经功能障碍的疾病,严重危害人类健康.近年来,基于质谱的蛋白质组学技术在神经退行性疾病的研究中得到了广泛应用.本文简要介绍了蛋白质组学在样品分离、多肽定量、质谱检测及生物标志物临床验证等方面的技术发展,并结合实例综述了基于质谱的蛋白质组学在神经退行性疾病生物标志物发现与验证中的研究进展.     

完整糖基化肽段的富集与质谱解析新技术研究进展

蛋白质糖基化是生物体内最重要的翻译后修饰之一,在蛋白质稳定性、细胞内和细胞间信号转导、激素活化或失活和免疫调节等生理过程和病理进程中发挥重要作用。而异常的蛋白质糖基化往往和多种疾病的发生发展密切相关,目前应用于临床检测的多种肿瘤生物标志物大多属于糖蛋白或者糖抗原。因此在组学层次系统分析蛋白质糖基化的变化对阐明生物体内糖基化修饰的调控机理和发现新型疾病标志物都非常重要。基于质谱的蛋白质组学技术为全面分析蛋白质及其修饰提供了有效的分析手段。在自下而上的蛋白质组学研究中,由于完整糖基化肽段同时存在性质各异的肽段骨架和糖链结构、糖肽的相对丰度和离子化效率较低以及糖基化修饰有高度异质性等特点,完整糖肽的分析比其他翻译后修饰更加困难。近年来,为了更全面、系统地分析蛋白质糖基化,研究人员发展了一些新技术,包括完整糖肽的富集技术、质谱的碎裂模式和数据采集模式、质谱数据的解析方法和定量策略等等,大力推进了该领域的研究水平,也为研究蛋白质糖基化相关的生物标志物提供了技术支持。该篇综述主要关注近年来基于质谱的糖蛋白质组学研究中的新进展,重点介绍针对完整N-和O-糖基化肽段的富集新技术和谱图解析新方法,并讨论其在肿瘤早期诊断方面的应用潜力。     

基于生物质谱的定量蛋白质组学研究进展

随着蛋白质组研究和生物质谱技术的发展,大规模的蛋白质组相对定量和绝对定量已经成为了解生命活动进程、疾病发生发展过程以及生物标志物筛选和验证的重要策略,并形成蛋白质组学研究领域的一个重要分支：定量蛋白质组学.综述了近年来定量蛋白质组学的研究进展,并对其中的关键技术进行讨论.     

基于ICP-MS的蛋白质定量方法*

蛋白质组学已经成为生命科学研究中最为活跃的领域之一。研究蛋白质的生物功能,不但需要高通量的鉴定蛋白质,还需要定量分析动态变化的蛋白质,即定量蛋白质组学研究。蛋白质的定量研究有助于发现新的生物功能,并可以用于疾病的预警和药物靶点的发现。现有的定量蛋白质组学研究主要利用同位素标记结合生物质谱（电喷雾电离质谱ESI-MS,基质辅助激光解吸电离质谱MALDI-MS）技术而实现。近年来电感耦合等离子体质谱（ICP-MS）作为ESI-MS和MALDI-MS的补充,越来越多地应用于蛋白质的定量分析,特别是蛋白质的绝对定量分析。ICP-MS是检测生物分子中痕量元素的理想工具,具有灵敏度高、动态范围广,不易受基体的影响等优点。本文将讨论基于ICP-MS的分析方法,及其在蛋白质定量分析和免疫分析中的部分成功应用。     

基于核酸适配体的生物标志物检测方法研究进展

核酸适配体是指能与特定靶分子结合的寡核苷酸链。因其具有易修饰、易合成和低免疫原性等特点,作为生物传感器的靶向元件,已广泛应用于各种生物标志物检测技术中。本文综述了近年来基于核酸适配体的生物标志物的检测技术在癌症、心血管疾病、阿尔兹海默症、抑郁症等多个疾病领域的诊断研究进展,列举核酸适配体新兴技术的创新应用,以期为生物标志物的检测提供新思路,也有望为相关疾病的早期诊断和治疗提供参考。     

蛋白质组学方法研究葡萄糖对人血清白蛋白的修饰作用

晚期糖基化终产物是一组由不同种类物质组成的复杂混合物,其与很多疾病相关,如糖尿病及其并发症、肾衰竭以及与老龄化相关等疾病.人血清白蛋白(HSA)是血清中含量最丰富的蛋白质.葡萄糖对血液中的HSA和其它蛋白质的结构具有修饰作用,发生糖基化反应.本文利用LC/MS联用技术,通过蛋白质组学方法,寻找差异肽段,研究糖基化HSA的修饰位点以及修饰种类,对于进一步寻找临床诊断糖尿病以及与老龄化相关疾病的肽类生物标志物具有重要意义.     

基于质谱的单细胞蛋白质组学分析方法及应用

细胞是生命体的最小组成单位,遗传及外部环境等因素使单细胞异质性广泛存在于众多生物体中。传统的生物学实验获得的结果多是大量细胞的平均测量值,因此在单细胞层面开展研究对于精确理解细胞的生长发育以及疾病的诊断与治疗至关重要。而作为重要的细胞和生命活动的执行者,蛋白质由于其不具备扩增特性,且种类繁多、丰度低、动态分布范围宽,与核酸等其他生物大分子相比,其单细胞组学研究相对滞后。而在所有的检测手段中,荧光检测以及电化学分析方法具有极高的灵敏度,但是囿于其研究通量有限,以及电化学活性依赖,很难成为普适性的单细胞蛋白质组学研究方法。质谱分析作为传统蛋白质组学中最为核心的研究技术,由于其高灵敏、高通量、结构信息丰富等特点,在单细胞蛋白质组学研究中独树一帜。该文综述了近年来基于质谱的单细胞蛋白质组学研究中的代表性方法,根据质谱分析前蛋白质分离方式的差异,将其分为基于毛细管电泳分离、液相色谱分离和无分离手段的直接检测3类方法,在介绍研究现状的同时对这些方法在细胞通量、蛋白质鉴定数目、灵敏度以及方法应用方面进行了总结与比较。最后,基于目前研究中面临的挑战以及发展趋势对基于质谱的单细胞蛋白质组学的研究前景进行了展望。     

基于多级质谱的蛋白质组定量新方法新技术进展

定量蛋白质组学已经成为蛋白质组学的一个重要分支,以生物质谱为核心的定量蛋白质组方法日益发展。按照定量所依据的质谱信号来源于一级质谱谱图还是多级质谱谱图可以将定量蛋白质组方法分为一级质谱定量和多级质谱定量。本文主要综述基于多级质谱的定量方法和技术进展,分析比较了这些方法的优缺点,并对基于多级质谱的定量方法发展进行了展望。     

福尔马林固定石蜡包埋组织切片蛋白质组分析方法与应用研究进展

福尔马林固定石蜡包埋(Formalin-fixed and paraffin-embedding,FFPE)是最常见的临床组织保存技术,FFPE组织具有标准化的制备流程、易于存储、标本量大、包含较完整的临床回顾性信息等特点,而成为疾病生物标志物发掘的良好载体。近年来,基于临床FFPE组织的特点,发展了系列蛋白质组学研究方法,包括样本制备、消解、分离到蛋白质质谱鉴定等多个领域,总体呈现出微量、高灵敏度、高通量的技术特点,并已成功用于肿瘤精准医学等临床蛋白质组研究。该综述将对FFPE组织切片样本的蛋白质组学方法,以及其在肿瘤研究中的应用进行概述,从而为临床精准医学蛋白质组学的研究提供借鉴思路。     

电化学生物传感应用于体外检测的研究

电化学生物传感技术以它独特的检测、分析方法以及在临床检测中潜在的应用,近年来受到研究者越来越多的关注。癌症生物标志物的早期检测能够使得患者在癌症发展至晚期前得到治疗,增加患者存活率。此外,生物标志物能够用于确定疾病的复发,以及患者在接受化疗、放疗和外科治疗之后的后续评估。本文主要论述了现有癌症生物标志物检测的设备和方法,并对这些方法的优点和不足作了简单的评述。另外,介绍了体外诊断设备的发展现状和电化学传感技术的特点,并对癌症早期的主要生物标志物进行了介绍,以及着重论述了电化学传感技术应用于临床靶向生物标志物检测的研究进展。此外,还展望了电化学生物传感技术未来的研究方向和发展趋势。从目前的的研究来看,电化学传感技术在体外诊断和临床检测癌症生物标志物等方面存在着较大的应用潜质,有望成为生物、医学、环境等领域重要的研究技术。     

Modified screen printed electrode for development of a highly sensitive label-free impedimetric immunosensor to detect amyloid beta peptides

Alzheimer's disease (AD) is a fatal neurodegenerative disease affecting approximately 26 million people world-wide, and the number is increasing as life expectancy increases. Since the only reliable diagnosis for the pathology is histochemical post-mortem examination, there is a rather urgent need for reliable, sensitive and quick detection techniques. Amyloid beta, being one of the established and widely accepted biomarkers of AD is a target biomolecule.     

Biomarkers for Alzheimer's Disease – An Overview

Alzheimer's disease (AD) is a complex neurodegenerative disorder with a significant global impact on public health. The emergence of atypical clinical phenotypes challenges traditional diagnostic approaches, necessitating a deeper exploration of biomarkers for accurate identification. The US Food and Drug Administration (FDA) classification of biomarkers and their integration into different stages of AD provide a structured framework for their application in research and clinical settings. Within the context of AD drug development, biomarkers are essential for participant selection, target engagement evaluation, and assessment of pathological hallmarks, including Aβ and tau protein abnormalities. The incorporation of nanoparticles with a biodegradable approach introduces innovative strategies to address the complexities of AD. This paper extensively discusses biomarkers associated with synaptic dysfunction, neuroinflammation, and glial activation, recognizing their significance in elucidating disease mechanisms. Common pathologies such as synuclein and TDP-43 further underscore the multifaceted nature of AD. Current biomarkers for AD diagnosis, encompassing cerebral spinal fluid (CSF) biomarkers and various imaging modalities, reflect the ongoing efforts to enhance early detection and monitoring. Intriguingly, novel biomarkers continue to emerge, offering promising avenues for improved understanding and intervention. Current review provides a comprehensive survey of biomarkers for AD, elucidating their diverse roles across different aspects of the disease. By highlighting their contributions to diagnosis, drug development, and mechanistic insights, this overview underscores the importance of biomarker research in the pursuit of effective AD management and treatment strategies.     

Recent Electrokinetic and Microfluidic Strategies for Detection of Amyloid Beta Peptide Biomarkers: Towards Molecular Diagnosis of Alzheimer's Disease

Among all neurodegenerative diseases, Alzheimer's Disease (AD) is the most prevalent worldwide, with a huge burden to the society and no efficient AD treatment so far. Continued efforts have been being made towards early and powerful diagnosis of AD, in the hope for a successful set of clinical trials and subsequently AD curative treatment. Towards this aim, detection and quantification of amyloid beta (Aβ) peptides in cerebrospinal fluid (CSF) and other biofluids, which are established and validated biomarkers for AD, have drawn attention of the scientific community and industry over almost two decades. In this work, an overview on our major contributions over 15 years to develop different electrokinetic and microfluidic strategies for Aβ peptides detection and quantification is reported. Accordingly, discussions and viewpoints on instrumental and methodological developments for microscale electrophoresis, microfluidic designs and immuno‐enrichment / assays on magnetic beads in microchannels for tracing Aβ peptides in CSF are given in this review.     

Blood-based Alzheimer's disease diagnosis using fluorescent peptide nanoparticle arrays

There is a critical need to diagnose and monitor the progression of Alzheimer’s disease(AD) using bloodbased biomarkers. At present, it is believed that tau biomarkers can be utilized to reliably detect AD.Multimodal techniques are highly sought after for AD diagnosis and progression monitoring. For this purpose, we developed a fluorescent peptide nanoparticles(f-PNPs) arrays that is capable of detecting multiple signals simultaneously. The concentration, aggregation stages, and Young’s modulus ...     

UPLC-TOF MS-Based Metabonomic Study on Coadministration of Huperzine A and Ligustrazine Phosphate for Treatment of Alzheimer’s Disease

This work was designed to assess the holistic efficacy of concurrent treatment with huperzine A (HA) and ligustrazine phosphate (LP) in the animal model, and to interpret the metabonomics characteristics of the poly-therapy in treating Alzheimer’s disease (AD). Metabolic profiling of the brain was performed using ultra performance liquid chromatography coupled to time-of-flight mass spectroscopy (UPLC-TOF MS). Principal component analysis and partial least squares discriminant analysis were utilized to classify and reveal the differences among the control, model and treatment groups. Ten potential biomarkers have been finally gained. The metabonomic results revealed that the coadministration of HA and LP might slow down the progression of AD through synergistic therapeutic efficacies, exhibiting better effect on amnesia compared with the mono-drug therapy. Importantly, the UPLC-TOF MS-based metabonomic approach would be a useful technique for the diagnosis of AD, the discovery of metabolic biomarkers and the evaluation of the treatment effects.

     

UPLC-TOF MS-Based Metabonomic Study on Coadministration of Huperzine A and Ligustrazine Phosphate for Treatment of Alzheimer��s Disease

This work was designed to assess the holistic efficacy of concurrent treatment with huperzine A (HA) and ligustrazine phosphate (LP) in the animal model, and to interpret the metabonomics characteristics of the poly-therapy in treating Alzheimer??s disease (AD). Metabolic profiling of the brain was performed using ultra performance liquid chromatography coupled to time-of-flight mass spectroscopy (UPLC-TOF MS). Principal component analysis and partial least squares discriminant analysis were utilized to classify and reveal the differences among the control, model and treatment groups. Ten potential biomarkers have been finally gained. The metabonomic results revealed that the coadministration of HA and LP might slow down the progression of AD through synergistic therapeutic efficacies, exhibiting better effect on amnesia compared with the mono-drug therapy. Importantly, the UPLC-TOF MS-based metabonomic approach would be a useful technique for the diagnosis of AD, the discovery of metabolic biomarkers and the evaluation of the treatment effects.     

From Chemistry to Translational Medicine: The Application of Proteomics to Cancer Biomarker Discovery and Diagnosis

The study of complex protein mixtures and their interactions in cells and tissues has been difficult due to the tedious process involved in their characterization and analysis. The recent emergence of fast‐evolving and state‐of‐the‐art proteomics methodologies has provided a rapid and scalable platform for understanding the comprehensive proteome profiles from complex whole tissues or cells of various biological sources. Therefore, proteomics has been increasingly valuable to examine real‐time changes in protein expression of various tissues or body fluids from patients with various diseases, especially cancer, resulting in the identification of clinically useful biomarkers for diagnosis, prognosis and disease staging. In this review, we focus on potential biomarkers for (1) Helicobacter pylori‐associated gastric cancer, (2) hepatocellular carcinoma (HCC), and (3) renal cell carcinoma (RCC). In addition to the conventional gel‐based proteomics (1‐D or 2‐D gels), we have utilized a more advanced proteomic approach by incorporating stable isotope dimethyl labelling and shotgun proteomics strategy in combination with nanoliquid chromatography and tandem mass spectrometry (nanoLC‐MS/MS) to better characterize the biomarkers in several cancer tissues. By establishing a high‐throughput proteomics platform based on multiple reaction monitoring (MRM), we have successfully detected and analyzed potential protein markers at low concentrations in various normal and tumor tissues. This platform not only highlights the utility of proteomics for biomarker discovery but also can be uniquely applied to disease‐oriented translational medicine for diagnosis of diverse types of cancers and other diseases.     

Application of NMR metabolomics to search for human disease biomarkers

Since antiquity, humans have used body fluids like saliva, urine and sweat for the diagnosis of diseases. The amount, color and smell of body fluids are still used in many traditional medical practices to evaluate an illness and make a diagnosis. The development and application of analytical methods for the detailed analysis of body fluids has led to the discovery of numerous disease biomarkers. Recently, mass spectrometry (MS), nuclear magnetic resonance spectroscopy (NMR), and multivariate statistical techniques have been incorporated into a multidisciplinary approach to profile changes in small molecules associated with the onset and progression of human diseases. The goal of these efforts is to identify metabolites that are uniquely correlated with a specific human disease in order to accurately diagnose and treat the malady. In this review we will discuss recent developments in sample preparation, experimental techniques, the identification and quantification of metabolites, and the chemometric tools used to search for biomarkers of human diseases using NMR.     

Lipidomic analysis and diagnosis of glioblastoma multiforme with rapid evaporative ionization mass spectrometry

Glioblastoma multiforme (GBM) is the most common and aggressive brain tumor in the central nervous system. GBM patients have a very low 5-year survival rate and most of them died within 1 year. Conventional histopathological examination for GBM diagnosis is complicated and time-consuming, which always blocks the development of more precise and effective treatments in resection operation. Rapid evaporative ionization mass spectrometry (REIMS) is a MS technique in clinical medicine research, which combines the common diathermy device with MS to acquire the lipid profiles of tissue specimens for lipidomic analysis and real-time tumor diagnosis. In this study, the REIMS method employing bipolar forceps was optimized and validated for high-throughput lipidomics and diagnosis of GBM for the first time. Total 42 lipid metabolites were tentatively identified and 12 out of 13 lipid biomarkers showed higher intensities in GBM, which were consistent with previous studies. After this, a statistic model was built with the lipidomic data for the diagnosis of GBM tumor in real-time. The diagnostic accuracy (94.74%), sensitivity (95.38%), and specificity (93.33%) were evaluated with histopathology validated brain tissue specimens that were not used in the training set. The proposed REIMS method for the lipidomic-analysis and diagnosis of GBM tumor provides a new direction for MS-based lipidomics and precision medicine and might be used to guide surgeons to precisely resect the GBM tissue and keep the normal brain tissue in operation.     

Molecularly imprinted monoliths: Recent advances in the selective recognition of biomacromolecules related biomarkers

Biomarkers are significant indicators to assist the early diagnosis of diseases and assess the therapeutic response. However, due to the low abundance of biomarkers in complex biological fluids, it is highly desirable to explore efficient techniques to attain their selective recognition and capture before the detection. Molecularly imprinted monoliths integrate the high selectivity of imprinted polymers and the rapid convective mass transport of monoliths, and as a result, are promising candidates to achieve the specific enrichment of biomarkers from complex samples. This review summarizes the various imprinting approaches for the preparation of molecularly imprinted monoliths. The state-of-art advances as an effective platform for applications in the selective capture of biomacromolecules related biomarkers were also outlined.