Antibody Repertoire Profiling Using Bacterial Display Identifies Reactivity Signatures of Celiac Disease
| Autor: | Joseph A. Murray, Bradley N. Spatola, Patrick S. Daugherty, Katri Kaukinen, Martin F. Kagnoff |
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| Rok vydání: | 2012 |
| Předmět: |
Adult
Male Tissue transglutaminase Peptide Sensitivity and Specificity Antibodies Article Analytical Chemistry Antigen-Antibody Reactions Antibody Repertoire Antigen Peptide Library Escherichia coli Humans Peptide library chemistry.chemical_classification Bacterial display biology Chemistry Antibody titer Middle Aged Flow Cytometry Molecular biology Celiac Disease biology.protein Female Antibody Cell Surface Display Techniques Peptides |
| Zdroj: | Analytical Chemistry. 85:1215-1222 |
| ISSN: | 1520-6882 0003-2700 |
| Popis: | A general strategy to identify serum antibody specificities associated with a given disease state, and peptide reagents for their detection was developed using bacterial display peptide libraries and multiparameter flow cytometry (MPFC). Using sera from patients with celiac disease (CD) (n = 45) or healthy subjects (n = 40), bacterial display libraries were screened for peptides that react specifically with antibodies from CD patients and not those from healthy patients. The libraries were screened for peptides that simultaneously cross-react with CD patient antibodies present in two separate patient groups labeled with spectrally distinct fluorophores, but do not react with unlabeled non-CD antibodies, thus affording a quantitative separation. A panel of six unique peptide sequences yielded 85% sensitivity and 91% specificity (AUC = 0.91) on a set of 60 samples not used for discovery, using leave-one-out cross-validation (LOOCV). Individual peptides were dissimilar with known CD specific antigens tissue transglutaminase (tTG) and deamidated gliadin, and classifier accuracy was independent of anti-tTG antibody titer. These results demonstrate that bacterial display/MPFC provides a highly effective tool for the unbiased discovery of disease-associated antibody specificities and peptide reagents for their detection that may have broad utility for diagnostic development. |
| Databáze: | OpenAIRE |
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