Identification of proteins directly from tissue: <i>in situ</i> tryptic digestions coupled with imaging mass spectrometry

Groseclose, M. Reid; Andersson, Malin; Hardesty, William M.; Caprioli, Richard M.

doi:10.1002/jms.1177

Cited by 353 publications

(415 citation statements)

References 33 publications

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“…Concerning MALDI-MSI, a lot of work has been done to improve the experimental workflow, such as optimization of sample preparation and matrix deposition [92], and much focus has been on the identification of the detected molecules. When small molecules are analyzed, the identification is rather straightforward where tandem mass spectrometry can be easily used, leading to the fragmentation of the detected compounds, allowing their unambiguous characterization.…”

Section: Maldi Mass Spectrometry Imagingmentioning

confidence: 99%

MALDI In-Source Decay, from Sequencing to Imaging

Debois

Smargiasso

Demeure

et al. 2012

Topics in Current Chemistry

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Matrix-assisted laser desorption/ionization (MALDI) is now a mature method allowing the identification and, more challenging, the quantification of biopolymers (proteins, nucleic acids, glycans, etc). MALDI spectra show mostly intact singly charged ions. To obtain fragments, the activation of singly charged precursors is necessary, but not efficient above 3.5 kDa, thus making MALDI MS/MS difficult for large species. In-source decay (ISD) is a prompt fragmentation reaction that can be induced thermally or by radicals. As fragments are formed in the source, precursor ions cannot be selected; however, the technique is not limited by the mass of the analyzed compounds and pseudo MS3 can be performed on intense fragments. The discovery of new matrices that enhance the ISD yield, combined with the high sensitivity of MALDI mass spectrometers, and software development, opens new perspectives. We first review the mechanisms involved in the ISD processes, then discuss ISD applications like top-down sequencing and post-translational modifications (PTMs) studies, and finally review MALDI-ISD tissue imaging applications.

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Section: Maldi Mass Spectrometry Imagingmentioning

confidence: 99%

MALDI In-Source Decay, from Sequencing to Imaging

Debois

Smargiasso

Demeure

et al. 2012

Topics in Current Chemistry

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“…This technology has progressed to the high-throughput proteomic profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections after ontissue tryptic digestion and to the top-down identification of most abundant proteins. This method was applied to FFPE-archived specimens in different tissue types, including lung cancer (21,22).…”

Section: Proteomic Analysis Of Invasive Tumorsmentioning

confidence: 99%

Advances in Proteomic Strategies toward the Early Detection of Lung Cancer

Hassanein¹,

Rahman²,

Chaurand³

et al. 2011

Proceedings of the American Thoracic Society

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TennesseeSince the advent of the new proteomics era more than a decade ago, large-scale studies of protein profiling have been exploited to identify the distinctive molecular signatures in a wide array of biological systems spanning areas of basic biological research, various disease states, and biomarker discovery directed toward therapeutic applications. Recent advances in protein separation and identification techniques have significantly improved proteomics approaches, leading to enhancement of the depth and breadth of proteome coverage. Proteomic signatures specific for invasive lung cancer and preinvasive lesions have begun to emerge. In this review we provide a critical assessment of the state of recent advances in proteomic approaches and the biological lessons they have yielded, with specific emphasis on the discovery of biomarker signatures for the early detection of lung cancer.Keywords: proteomics; biomarker; early detection; lung cancer CHALLENGES IN LUNG CANCER RESEARCHLung cancer is the deadliest cancer in the United States and worldwide, accounting for 15% of all cancer incidence and 29% of all cancer deaths, with a 5-year survival rate of only 15% (1, 2). Lung cancer represents a spectrum of diseases with tremendous heterogeneity at the pathological and molecular levels (3-6) that is strongly associated with smoking as a risk factor. With about 20% of the United States adult population smoking and 1 billion smokers worldwide, it was estimated that in 2009 lung cancer claimed more lives than breast, prostate, colon, liver, kidney, and melanoma cancers combined (2, 7). Despite the recent improvements of bronchoscopic and surgical techniques as well as advances in chemotherapy and radiation therapy treatments, attempts to improve patient outcomes are faced with immense challenges. Several noninvasive detection technologies have been investigated. Imaging techniques, such as chest radiography, lowdose spiral computed tomography, sputum cytology, and molecular biomarkers in various biological samples, have been tested for their diagnostic value for early detection of lung cancer (8, 9). Although these tests vary in their sensitivity and specificity, only low-dose chest computed tomography was shown to reduce lung cancer-specific mortality (10-12). This encouraging finding calls for new molecular strategies to address the noninvasive diagnostic and risk assessment for lung cancer. These molecular strategies will have to demonstrate clinical utility and may complement currently tested strategies. NATURAL HISTORY OF LUNG CANCER PROGRESSION AND A WINDOW FOR EARLY DETECTIONLung cancer can be considered to result from a long history of repeated airway damage and repair cycles. Although clinically addressed at the time of diagnosis, the disease process develops for months and years before affecting patients' lives. This rather long disease process (Figure 1) represents a window of opportunity where the intervention should take place with the aim of preventing the development of disease (e.g., primary preven...

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“…For example, enzymatic digestion using trypsin was carried out in an ordered array of microspots across sections of rat brain with subsequent matrix application directly onto the digested spots [10]. The spatial selectivity of the digestion was demonstrated where multiple unique digested peptides from one brain section display the same distribution as the intact parent protein in a serial section.…”

Section: Applicationsmentioning

confidence: 99%