Abstract
Recently, attempts to reveal the structures of autoantibodies comprehensively using improved proteogenomics technology, have become popular. This technology identifies peptides in highly purified antibodies by using an Orbitrap device to compare spectra from liquid chromatography–tandem mass spectrometry against a cDNA database obtained through next-generation sequencing. In this study, we first analyzed granulocyte-macrophage colony-stimulating factor (GM-CSF) autoantibodies in a patient with autoimmune pulmonary alveolar proteinosis, using the trapped ion mobility spectrometry coupled with quadrupole time-of-flight (TIMS-TOF) instrument. The TIMS-TOF instrument identified peptides that partially matched sequences in up to 156 out of 162 cDNA clones. Complementarity-determining region 3 (CDR3) was fully and partially detected in nine and 132 clones, respectively. Moreover, we confirmed one unique framework region 4 (FR4) and at least three unique across CDR3 to FR4 peptides via de novo peptide sequencing. This new technology may thus permit the comprehensive identification of autoantibody structure.
Highlights
Attempts to reveal the structures of autoantibodies comprehensively using improved proteogenomics technology, have become popular
Our previous study have reported that circulating granulocyte-macrophage colony-stimulating factor (GM-CSF) autoreactive B cells (GMARBs) producible of GM-CSF specific antibodies (GMAbs) by Epstein-Barr virus transformation are consistently detected in healthy individuals and patients with aPAP6
From the total RNA, full-length cDNA with the SMARTer-oligo sequence at 5′ end was subjected to polymerase chain reaction (PCR) to generate amplicons for variable regions of the IgG heavy chain (IgG-HV)
Summary
Attempts to reveal the structures of autoantibodies comprehensively using improved proteogenomics technology, have become popular. We first analyzed granulocyte-macrophage colony-stimulating factor (GM-CSF) autoantibodies in a patient with autoimmune pulmonary alveolar proteinosis, using the trapped ion mobility spectrometry coupled with quadrupole time-of-flight (TIMS-TOF) instrument. We confirmed one unique framework region 4 (FR4) and at least three unique across CDR3 to FR4 peptides via de novo peptide sequencing This new technology may permit the comprehensive identification of autoantibody structure. Our previous study have reported that circulating GM-CSF autoreactive B cells (GMARBs) producible of GMAb by Epstein-Barr virus transformation are consistently detected in healthy individuals and patients with aPAP6. These are potential precursor cells for GMAb-producing plasma cells and B cells
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
