NISA: Novel approaches for in situ analysis of biomolecules

NISA：生物分子原位分析的新方法

• 批准号: EP/L023490/1
• 负责人: Helen Cooper
• 金额: $ 189.16万
• 依托单位: University of Birmingham
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FL023490%2F1
• 关键词: NISA; Novel; approaches; situ; analysis

The aim of the research is to develop novel approaches for the analysis of biomolecules, and in particular proteins, directly from their natural (or actual) environment, i.e., to develop approaches for in situ biomolecular analysis. Proteins are the work-horses of the cell and perform all the functions required for life. They also find uses as therapeutics and in consumer products. To gain insight into the various and specific roles of proteins in life processes, or to determine the therapeutic efficacy of protein drugs, or to establish the environmental fate of protein additives in consumer products, it is necessary to be able to analyse proteins at a molecular level. Mass spectrometry, in which ionised molecules are characterised according to their mass-to-charge, is ideally suited to this challenge, offering high sensitivity, broad specificity (all molecules have a mass), and the capability for chemical structure elucidation. The ultimate goal is to link molecular analysis directly to molecular environment. Much like a forensics officer tasked with determining the presence of an illicit substance, there is much greater reliability and credibility afforded to an analysis performed at the scene of the crime than to one performed following removal of the sample to a separate location and alternative surroundings.Growing evidence suggests in situ protein analysis has groundbreaking roles to play in biomarker discovery, diagnosis & early detection of disease, targeting therapeutics (personalised medicine) and assessment of therapeutic efficacy. The benefits of in situ protein analysis can be illustrated by considering a thin tissue section through a drug-treated tumour. In principle, in situ analysis would inform on drug-target interactions (i.e., is the drug binding to the correct protein?). Moreover, with in situ protein analysis the capacity for artefact introduction as a result of sample preparation (e.g., application of a matrix) or sample damage is eliminated.Nevertheless, a number of challenges exist. Proteins are large molecules associated with a vast array of chemical modifications, and which form loosely-bound complexes with themselves, other proteins and other molecule types. It is not only their chemical structure but also their overall 3-D structure which dictate their function. Other molecular classes that are hugely important in biological processes also have an intricate relationship with proteins. Any in situ mass spectrometry approach needs to be able to meet these analyte-driven challenges, i.e., it must be capable of (a) measuring proteins and characterising any modifications, (b) detecting protein complexes and determining their constituents, (c) providing information on 3-D structure, and (d) detecting other relevant molecular classes. Moreover, there are technique-driven challenges for in situ analysis including inherently high sample complexity and wide ranging concentrations, and opportunities for quantitation.The research will meet these challenges by developing a newly emerging in situ approach, liquid extraction surface analysis mass spectrometry, in combination with two complementary types of ion mobility spectrometry (which can either provide information on 3-D structure, or separate ionised molecules in the mass spectrometer on the basis of their 3-D shape) and a structural elucidation strategy known as electron-mediated dissociation mass spectrometry.The research will be undertaken primarily at the University of Birmingham in the Advanced Mass Spectrometry Facility in the School of Biosciences and the School of Chemistry mass spectrometry facility. The programme involves a number of academic and industrial collaborators and additional research will be carried out during scientific visits to National Physical Laboratory (NPL), Thermo Fisher Scientific, Waters, Owlstone, Florida State University, Texas A&M University and Université d'Aix-Marseille.

这项研究的目的是开发新的方法来分析生物分子，尤其是蛋白质，直接从其自然（或实际）环境（即，即开发原位生物分子分析的方法）。蛋白质是细胞的工作马，并执行生命所需的所有功能。他们还发现用途作为治疗和消费产品。为了深入了解蛋白质在生命过程中的各种和特定的作用，或确定蛋白质药物的治疗有效性，或在消费产品中建立蛋白质添加剂的环境命运，有必要能够以分子水平分析蛋白质。质谱法根据其质量到电荷来表征离子化的分子，非常适合这一挑战，具有较高的灵敏度，广泛的特异性（所有分子均具有质量）以及化学结构阐明的能力。最终目标是将分子分析直接与分子环境联系起来。就像一名法医官员决定存在非法物质一样，对在犯罪现场进行的分析提供的可靠性和可信度比将样本移至单独的位置和替代环境后进行的分析要比进行的分析要大得多。越来越多的证据表明，原位蛋白分析在生物标志物发现，诊断和早期发现疾病，靶向治疗（个性化医学）和治疗评估中起着突破性的作用。原位蛋白分析的好处可以通过考虑通过药物治疗的肿瘤进行薄的组织部分来说明。原则上，原位分析将告知药物目标相互作用（即药物与正确的蛋白质结合？）。此外，通过原位蛋白分析，消除了样品制备（例如，矩阵的应用）或样品损伤的导致人工伪像引入的能力。尚无许多挑战。蛋白质是与大量化学修饰相关的大分子，它们与自身，其他蛋白质和其他分子类型形成了松散结合的复合物。决定其功能的不仅是他们的化学结构，而且是他们的总体3-D结构。在生物过程中非常重要的其他分子类也与蛋白质具有复杂的关系。任何原位质谱方法都需要能够应对这些分析驱动的挑战，即，它必须能够（a）测量蛋白质并表征任何修饰，（b）检测蛋白质复合物并确定其构建体，（c）提供有关3-D结构的信息，以及（d）检测其他相关分子类别。 Moreover, there are technique-driven challenges for in situ analysis including inherently high sample complexity and wide ranging concentrations, and opportunities for quantitation.The research will meet these challenges by developing a newly emerging in situ approach, liquid extraction surface analysis mass spectrometry, in combination with two complete types of ion mobility spectrometry (which can either provide information on 3-D structure, or separate ionised molecules in the mass spectrometer on the basis of their 3-D形状）和一种称为电子介导的解离质谱法的结构阐明策略。该研究将主要在伯明翰大学的生物科学学院和化学质谱学院的高级质谱学设施中进行。该计划涉及许多学术和工业合作者，在科学访问国家物理实验室（NPL），Thermo Fisher Scientific，Waters，Waters，Owlstone，Florida State University，Texas A＆M University和Universition d'Aix-Marseille期间将进行其他研究。

项目成果

Separation of cis and trans Isomers of Polyproline by FAIMS Mass Spectrometry.

• DOI: 10.1007/s13361-016-1482-1
• 发表时间: 2016-12
• 期刊: JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
• 影响因子: 3.2
• 作者: Creese, Andrew J.;Cooper, Helen J.
• 通讯作者: Cooper, Helen J.

Native mass spectrometry imaging of intact proteins and protein complexes in thin tissue sections

• DOI: 10.1016/j.ijms.2017.10.009
• 发表时间: 2019-03-01
• 期刊: INTERNATIONAL JOURNAL OF MASS SPECTROMETRY
• 影响因子: 1.8
• 作者: Griffiths, Rian L.;Sisley, Emma K.;Cooper, Helen J.
• 通讯作者: Cooper, Helen J.

To What Extent is FAIMS Beneficial in the Analysis of Proteins?

• DOI: 10.1007/s13361-015-1326-4
• 发表时间: 2016-04
• 期刊: Journal of the American Society for Mass Spectrometry
• 影响因子: 3.2
• 作者: Cooper HJ

Identification of Phosphorylation Sites Altering Pollen Soluble Inorganic Pyrophosphatase Activity

• DOI: 10.1104/pp.16.01450
• 发表时间: 2017-03-01
• 期刊: PLANT PHYSIOLOGY
• 影响因子: 7.4
• 作者: Eaves, Deborah J.;Haque, Tamanna;Franklin-Tong, Vernonica E.
• 通讯作者: Franklin-Tong, Vernonica E.