Proteomics

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Proteomic identification of anti-tetanus toxoid antibodies from human serum (HD1)

ABSTRACT: Proteomic identification and characterization of antibodies comprising the serological response to antigen can provide unique insight into the functional dynamics of adaptive immunity. We have developed a novel method to overcome the technical challenges which previously limited the direct analysis of immunoglobulin proteins in serum, as demonstrated by the identification of human anti-tetanus toxoid (TT) immunoglobulin G (IgG) proteins following booster vaccination. We analyzed the serum IgG repertoire across four time-points corresponding to pre-vaccination, 7 days, 3 months, and 9 months post vaccination. Antigen-specific antibodies were affinity purified against immobilized TT protein and sequenced by bottom-up nanoLC-MS/MS. Interpretation of mass spectra required a custom reference database of IgG heavy and light chain variable sequences determined by NextGen RNA sequencing of the donor's circulating plasmablasts and memory B cells following booster vaccination.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Serum

SUBMITTER: Daniel Boutz  

LAB HEAD: Edward Marcotte

PROVIDER: PXD000917 | Pride | 2014-04-24

REPOSITORIES: Pride

Json Xml

Dataset's files

Source:
Action DRS
TT_HD1_ConJ_reference_database.fasta Fasta
TT_HD1_D000a.raw Raw
TT_HD1_D000a_ConJ.mgf Mgf
TT_HD1_D000a_ConJ.pep.xml Pepxml
TT_HD1_D000a_ShuffleDB.mgf Mgf
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Publications

Proteomic identification of monoclonal antibodies from serum.

Boutz Daniel R DR   Horton Andrew P AP   Wine Yariv Y   Lavinder Jason J JJ   Georgiou George G   Marcotte Edward M EM  

Analytical chemistry 20140501 10

Characterizing the in vivo dynamics of the polyclonal antibody repertoire in serum, such as that which might arise in response to stimulation with an antigen, is difficult due to the presence of many highly similar immunoglobulin proteins, each specified by distinct B lymphocytes. These challenges have precluded the use of conventional mass spectrometry for antibody identification based on peptide mass spectral matches to a genomic reference database. Recently, progress has been made using botto  ...[more]

PMID: 24684310

Publication: 1/2

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