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面向临床执业医师的蛋白质组学方法与应用

Proteomics methods and applications for the practicing clinician.

作者信息

Reisdorph Nichole A, Reisdorph Richard, Bowler Russell, Broccardo Carolyn

机构信息

Department of Immunology, National Jewish Health, Denver, Colorado 80206, USA.

出版信息

Ann Allergy Asthma Immunol. 2009 Jun;102(6):523-9. doi: 10.1016/S1081-1206(10)60128-7.

Abstract

OBJECTIVE

To describe clinical proteomics from discovery techniques and their limitations, to applications in allergy, asthma, and immunology, and finally to how proteomics can be integrated into clinical practice.

DATA SOURCES

Despite many inherent challenges, proteomics-based methods have become a powerful and popular means of profiling clinical samples for the purpose of biomarker discovery. Although several strategies exist, clinical proteomics for the purpose of biomarker discovery generally focuses on 1 of 3 basic workflows: (1) 2-dimensional gel electrophoresis to quantitate relative protein levels followed by mass spectrometry (MS) to identify proteins of interest, (2) non-gel-based methods that rely on liquid chromatography MS (LCMS) for both quantitation and identification of proteins, and (3) protein profiling methods that do not directly result in the identification of proteins but rather generate "fingerprints" that are compared among individuals or samples.

STUDY SELECTION

Regardless of the strategy being pursued, a few general experimental steps are followed that will be expounded on in the text. These proteomics techniques have been applied to discover new biomarkers in biofluids and tissues from individuals with a variety of conditions, including allergy, asthma, atopic dermatitis, inflammatory diseases, chronic obstructive pulmonary disease, and other lung diseases.

RESULTS

After biomarker discovery, LCMS-based proteomics offers several advantages over traditional antibody-based clinical assays, including greater specificity, cost- and time-effectiveness, and the potential to multiplex up to hundreds of peptides in a single assay.

CONCLUSION

With many guidelines now in place and model studies on which to design future experiments, there is reason to be optimistic that candidate protein biomarkers will be discovered using proteomics and translated into clinical assays.

摘要

目的

描述从发现技术及其局限性到临床蛋白质组学在过敏、哮喘和免疫学中的应用,最后阐述蛋白质组学如何融入临床实践。

数据来源

尽管存在许多内在挑战,但基于蛋白质组学的方法已成为用于生物标志物发现的强大且流行的临床样本分析手段。虽然存在几种策略,但用于生物标志物发现的临床蛋白质组学通常聚焦于3种基本工作流程中的一种:(1)二维凝胶电泳定量相对蛋白质水平,随后通过质谱(MS)鉴定感兴趣的蛋白质;(2)基于非凝胶的方法,依赖液相色谱 - 质谱联用(LCMS)进行蛋白质定量和鉴定;(3)蛋白质谱分析方法,该方法不会直接鉴定蛋白质,而是生成可在个体或样本之间进行比较的“指纹图谱”。

研究选择

无论采用何种策略,都遵循一些通用的实验步骤,本文将对此进行阐述。这些蛋白质组学技术已应用于从患有多种疾病(包括过敏、哮喘、特应性皮炎、炎症性疾病、慢性阻塞性肺疾病和其他肺部疾病)的个体的生物流体和组织中发现新的生物标志物。

结果

在发现生物标志物后,基于LCMS的蛋白质组学相对于传统的基于抗体的临床检测具有几个优势,包括更高的特异性、成本效益和时间效益,以及在单次检测中对多达数百种肽进行多重分析的潜力。

结论

由于现在有许多指南以及可用于设计未来实验的模型研究,因此有理由乐观地认为,将通过蛋白质组学发现候选蛋白质生物标志物并将其转化为临床检测方法。

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