Fast Photochemical Oxidation of Proteins for Epitope Mapping
The growing use of monoclonal antibodies as therapeutics underscores the importance of epitope mapping as an essential step in characterizing antibody–antigen complexes. The use of protein footprinting coupled with mass spectrometry, which is emerging as a tool in structural biology, offers opportun...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2011-10, Vol.83 (20), p.7657-7661 |
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| container_title | Analytical chemistry (Washington) |
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| creator | Jones, Lisa M B. Sperry, Justin A. Carroll, James Gross, Michael L |
| description | The growing use of monoclonal antibodies as therapeutics underscores the importance of epitope mapping as an essential step in characterizing antibody–antigen complexes. The use of protein footprinting coupled with mass spectrometry, which is emerging as a tool in structural biology, offers opportunities to map antibody-binding regions of antigens. We report here the use of footprinting via fast photochemical oxidation of proteins (FPOP) with OH radicals to characterize the epitope of the serine protease thrombin. The data correlate well with previously published results that determined the epitope of thrombin. This study marks the first time oxidative labeling has been used for epitope mapping. |
| doi_str_mv | 10.1021/ac2007366 |
| format | Article |
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Sperry, Justin</creatorcontrib><creatorcontrib>A. Carroll, James</creatorcontrib><creatorcontrib>Gross, Michael L</creatorcontrib><title>Fast Photochemical Oxidation of Proteins for Epitope Mapping</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>The growing use of monoclonal antibodies as therapeutics underscores the importance of epitope mapping as an essential step in characterizing antibody–antigen complexes. The use of protein footprinting coupled with mass spectrometry, which is emerging as a tool in structural biology, offers opportunities to map antibody-binding regions of antigens. We report here the use of footprinting via fast photochemical oxidation of proteins (FPOP) with OH radicals to characterize the epitope of the serine protease thrombin. The data correlate well with previously published results that determined the epitope of thrombin. 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Psychology</subject><subject>Fundamental immunology</subject><subject>Hydroxyl Radical - chemistry</subject><subject>Mass Spectrometry</subject><subject>Molecular immunology</subject><subject>Oxidation</subject><subject>Oxidation-Reduction</subject><subject>Proteins</subject><subject>Spectrometric and optical methods</subject><subject>Thrombin - immunology</subject><subject>Thrombin - metabolism</subject><subject>Trypsin - metabolism</subject><issn>0003-2700</issn><issn>1520-6882</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNplkU-LFDEQxYMo7jh68AtII4h4aK1KJkkHRJBlV4WV3YOeQ3Um2cnS02mTHtFvb5YdZ_xzCkV-vHr1HmNPEV4jcHxDjgNoodQ9tkDJoVVdx--zBQCIlmuAE_aolBsARED1kJ1w7MzKGLVgb8-pzM3VJs3Jbfw2Ohqayx9xTXNMY5NCc5XT7ONYmpByczbFOU2--UzTFMfrx-xBoKH4J_t3yb6en305_dheXH74dPr-oqWVwrn1DoQBrvvQYZC6F5pTvxZBGx2kQuXRSEXKgZYr7HrhCao7QiBZJ23Ekr270512_davnR_nTIOdctxS_mkTRfv3zxg39jp9twKNkBKrwMu9QE7fdr7MdhuL88NAo0-7YjvTSdAab8nn_5A3aZfHep01gCutVc10yV7dQS6nUrIPBysI9rYRe2ikss_-9H4gf1dQgRd7gEpNP2QaXSxHrkbIBYojR64cTf2_8BeOwp0H</recordid><startdate>20111015</startdate><enddate>20111015</enddate><creator>Jones, Lisa M</creator><creator>B. 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| subjects | Analytical chemistry Antibodies, Monoclonal - immunology Antigen-Antibody Complex Antigens Biological and medical sciences Chemistry Chromatography, High Pressure Liquid Correlation analysis Epitope Mapping - methods Exact sciences and technology Fundamental and applied biological sciences. Psychology Fundamental immunology Hydroxyl Radical - chemistry Mass Spectrometry Molecular immunology Oxidation Oxidation-Reduction Proteins Spectrometric and optical methods Thrombin - immunology Thrombin - metabolism Trypsin - metabolism |
| title | Fast Photochemical Oxidation of Proteins for Epitope Mapping |
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