J. D. Meutter, Kheiro-Mouna Derfoufi, E. Goormaghtigh
{"title":"蛋白质微阵列的FTIR成像分析","authors":"J. D. Meutter, Kheiro-Mouna Derfoufi, E. Goormaghtigh","doi":"10.3233/BSI-160137","DOIUrl":null,"url":null,"abstract":"BACKGROUND: Proteins are sensitive to environmental conditions. Whether they are produced for therapeutic purposes or for fundamental research, the integrity of their structure and post-traductional modifications are key issues. Measuring glycosylation or phosphorylation level as well as their secondary structure most often rely on complex and indirect experiments. Infrared spectroscopy presents a series of advantages related to its multivariate character. There is a lack of high-throughput methods able to analyse these parameters. OBJECTIVE: In this paper we attempted to combine protein microarrays and infrared imaging for high throughput analysis of proteins. METHODS: A protein microarrayer was used to produce protein microarrays on BaF2 slides transparent in the mid-infrared. Spot density was about 25 spots/mm 2 . A 128 × 128 focal plane array infrared detector was used to record images of the protein microarrays. RESULTS: We show that 100 µm diameter spot are easily analyzed. Spots obtained with low protein concentrations, resulting in an average of a single protein monolayer (ca 3 fg/µm 2 for a 66 kDa protein) provided good quality spectra. CONCLUSIONS: Infrared imaging is a label free, high throughput method, able to analyse protein microarrays and to take advantage from the wide information available in the infrared spectra.","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.3000,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-160137","citationCount":"14","resultStr":"{\"title\":\"Analysis of protein microarrays by FTIR imaging\",\"authors\":\"J. D. Meutter, Kheiro-Mouna Derfoufi, E. Goormaghtigh\",\"doi\":\"10.3233/BSI-160137\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"BACKGROUND: Proteins are sensitive to environmental conditions. Whether they are produced for therapeutic purposes or for fundamental research, the integrity of their structure and post-traductional modifications are key issues. Measuring glycosylation or phosphorylation level as well as their secondary structure most often rely on complex and indirect experiments. Infrared spectroscopy presents a series of advantages related to its multivariate character. There is a lack of high-throughput methods able to analyse these parameters. OBJECTIVE: In this paper we attempted to combine protein microarrays and infrared imaging for high throughput analysis of proteins. METHODS: A protein microarrayer was used to produce protein microarrays on BaF2 slides transparent in the mid-infrared. Spot density was about 25 spots/mm 2 . A 128 × 128 focal plane array infrared detector was used to record images of the protein microarrays. RESULTS: We show that 100 µm diameter spot are easily analyzed. Spots obtained with low protein concentrations, resulting in an average of a single protein monolayer (ca 3 fg/µm 2 for a 66 kDa protein) provided good quality spectra. CONCLUSIONS: Infrared imaging is a label free, high throughput method, able to analyse protein microarrays and to take advantage from the wide information available in the infrared spectra.\",\"PeriodicalId\":44239,\"journal\":{\"name\":\"Biomedical Spectroscopy and Imaging\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.3000,\"publicationDate\":\"2016-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.3233/BSI-160137\",\"citationCount\":\"14\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biomedical Spectroscopy and Imaging\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.3233/BSI-160137\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"SPECTROSCOPY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biomedical Spectroscopy and Imaging","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.3233/BSI-160137","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"SPECTROSCOPY","Score":null,"Total":0}
BACKGROUND: Proteins are sensitive to environmental conditions. Whether they are produced for therapeutic purposes or for fundamental research, the integrity of their structure and post-traductional modifications are key issues. Measuring glycosylation or phosphorylation level as well as their secondary structure most often rely on complex and indirect experiments. Infrared spectroscopy presents a series of advantages related to its multivariate character. There is a lack of high-throughput methods able to analyse these parameters. OBJECTIVE: In this paper we attempted to combine protein microarrays and infrared imaging for high throughput analysis of proteins. METHODS: A protein microarrayer was used to produce protein microarrays on BaF2 slides transparent in the mid-infrared. Spot density was about 25 spots/mm 2 . A 128 × 128 focal plane array infrared detector was used to record images of the protein microarrays. RESULTS: We show that 100 µm diameter spot are easily analyzed. Spots obtained with low protein concentrations, resulting in an average of a single protein monolayer (ca 3 fg/µm 2 for a 66 kDa protein) provided good quality spectra. CONCLUSIONS: Infrared imaging is a label free, high throughput method, able to analyse protein microarrays and to take advantage from the wide information available in the infrared spectra.
期刊介绍:
Biomedical Spectroscopy and Imaging (BSI) is a multidisciplinary journal devoted to the timely publication of basic and applied research that uses spectroscopic and imaging techniques in different areas of life science including biology, biochemistry, biotechnology, bionanotechnology, environmental science, food science, pharmaceutical science, physiology and medicine. Scientists are encouraged to submit their work for publication in the form of original articles, brief communications, rapid communications, reviews and mini-reviews. Techniques covered include, but are not limited, to the following: • Vibrational Spectroscopy (Infrared, Raman, Teraherz) • Circular Dichroism Spectroscopy • Magnetic Resonance Spectroscopy (NMR, ESR) • UV-vis Spectroscopy • Mössbauer Spectroscopy • X-ray Spectroscopy (Absorption, Emission, Photoelectron, Fluorescence) • Neutron Spectroscopy • Mass Spectroscopy • Fluorescence Spectroscopy • X-ray and Neutron Scattering • Differential Scanning Calorimetry • Atomic Force Microscopy • Surface Plasmon Resonance • Magnetic Resonance Imaging • X-ray Imaging • Electron Imaging • Neutron Imaging • Raman Imaging • Infrared Imaging • Terahertz Imaging • Fluorescence Imaging • Near-infrared spectroscopy.