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dc.contributor.authorDíez, Paula-
dc.contributor.authorIbarrola, Nieves-
dc.contributor.authorDégano, Rosa M.-
dc.contributor.authorLécrevisse, Quentin-
dc.contributor.authorRodríguez-Caballero, Arancha-
dc.contributor.authorCriado, Ignacio-
dc.contributor.authorNieto, Wendy G.-
dc.contributor.authorGóngora, Rafael-
dc.contributor.authorGonzález, Marcos-
dc.contributor.authorAlmeida, Julia-
dc.contributor.authorOrfao, Alberto-
dc.contributor.authorFuentes, Manuel-
dc.identifierdoi: 10.18632/oncotarget.17076-
dc.identifiere-issn: 1949-2553-
dc.identifier.citationOncotarget 8(26): 42836-42846 (2017)-
dc.description.abstractA wide variety of immunoglobulins (Ig) is produced by the immune system thanks to different mechanisms (V(D)J recombination, somatic hypermutation, and antigen selection). The profiling of Ig sequences (at both DNA and peptide levels) are of great relevance to developing targeted vaccines or treatments for specific diseases or infections. Thus, genomics and proteomics techniques (such as Next- Generation Sequencing (NGS) and mass spectrometry (MS)) have notably increased the knowledge in Ig sequencing and serum Ig peptide profiling in a high-throughput manner. However, the peptide characterization of membrane-bound Ig (e.g., B-cell receptors, BCR) is still a challenge mainly due to the poor recovery of mentioned Ig. Herein, we have evaluated three different sample processing methods for peptide sequencing of BCR belonging to chronic lymphocytic leukemia (CLL) B cells identifying up to 426 different peptide sequences (MS/MS data are available via ProteomeXchange with identifier PXD004466). Moreover, as a consequence of the results here obtained, recommended guidelines have been described for BCR-sequencing of B-CLL samples by MS approaches. For this purpose, an in-house algorithm has been designed and developed to compare the MS/MS results with those obtained by molecular biology in order to integrate both proteomics and genomics results and establish the steps to follow when sequencing membrane-bound Ig by MS/MS.-
dc.description.sponsorshipWe gratefully acknowledge financial support from the Spanish Health Institute Carlos III (ISCIII) for the grants: FIS PI11/02114 and FIS PI114/01538. We also acknowledge Fondos FEDER (EU) and Junta Castilla León (grant BIO/SA07/15). This work has been also sponsored by Fundación Solórzano (FS/23-2015). The Proteomics Unit belongs to ProteoRed, PRB2-ISCIII, supported by grant PT13/0001, of the PE I+D+I 2013-2016, funded by ISCIII and FEDER. The authors would like to thank all the clinicians and technicians in the Cytometry and Cell Purification Services of the University of Salamanca, the Spanish National DNA Bank (Banco Nacional de DNA Carlos III, University of Salamanca) and the Genomic Unit of Cancer Research Centre (IBMCC, USAL-CSIC) for their support in the data collection for the preparation of this manuscript. P.D. is supported by a JCYL-EDU/346/2013 Ph.D. scholarship.-
dc.publisherImpact Journals-
dc.relation.isversionofPublisher's version-
dc.subjectPeptide sequencing-
dc.subjectMass spectrometry-
dc.subjectB-cell receptor-
dc.subjectChronic lymphocytic leukemia-
dc.titleA systematic approach for peptide characterization of B-cell receptor in chronic lymphocytic leukemia cells-
dc.description.versionPeer Reviewed-
dc.contributor.funderEuropean Commission-
dc.contributor.funderJunta de Castilla y León-
dc.contributor.funderInstituto de Salud Carlos III-
dc.contributor.funderFundación Memoria de D. Samuel Solorzano Barruso-
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