کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
7636153 | 1494771 | 2014 | 5 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Accurate quantitation of MHC-bound peptides by application of isotopically labeled peptide MHC complexes
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کلمات کلیدی
AQUAHSCTpMHCEBV-LCLMiHAHuman leukocyte antigen - آنتی ژن لوسکسی انسانMinor histocompatibility antigens - آنتی ژن های هیستوپاتولوژی کوچکHLA - آنتیژن گلبول سفید انسانیimmunoaffinity purification - تصفیه ایمنی آلایندهphycoerythrin - فایکوئیریدینAbsolute quantitation - مقدار مطلقHematopoietic stem cell transplantation - پیوند مغز استخوان
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Knowledge of the accurate copy number of HLA class I presented ligands is important in fundamental and clinical immunology. Currently, the best copy number determinations are based on mass spectrometry, employing single reaction monitoring (SRM) in combination with a known amount of isotopically labeled peptide. The major drawback of this approach is that the losses during sample pretreatment, i.e. immunopurification and filtration steps, are not well defined and must, therefore, be estimated. In addition, such losses can vary for individual peptides. Therefore, we developed a new approach in which isotopically labeled peptide-MHC monomers (hpMHC) are prepared and added directly after cell lysis, i.e. before the usual sample processing. Using this approach, all losses during sample processing can be accounted for and allows accurate determination of specific MHC class I-presented ligands. Our study pinpoints the immunopurification step as the origin of the rather extreme losses during sample pretreatment and offers a solution to account for these losses. Obviously, this has important implications for accurate HLA-ligand quantitation. The strategy presented here can be used to obtain a reliable view of epitope copy number and thus allows improvement of vaccine design and strategies for immunotherapy.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Proteomics - Volume 109, 23 September 2014, Pages 240-244
Journal: Journal of Proteomics - Volume 109, 23 September 2014, Pages 240-244
نویسندگان
Chopie Hassan, Michel G.D. Kester, Gideon Oudgenoeg, Arnoud H. de Ru, George M.C. Janssen, Jan W. Drijfhout, Robbert M. Spaapen, Connie R. Jiménez, Mirjam H.M. Heemskerk, J.H. Frederik Falkenburg, Peter A. van Veelen,