Intact Transition Epitope Mapping – Serological Inspection by Epitope EXtraction (ITEM – SIX)

ABSTRACT: Precision medicine requests accurate serological inspections to precisely stratify patients for targeted treatment. Intact transition epitope mapping analysis proved seroconversion of a model organism's serum when spiked with a monoclonal murine anti-ovalbumin antibody with epitope-resolution. Isolation of the IgG fraction from blood serum applied two consecutive protein precipitation steps followed by ultrafiltration and resulted in an ESI-MS analysis-ready IgG preparation. For epitope mapping by epitope extraction, the ovalbumin antigen was digested with trypsin. After desalting, the peptide mixture was added to the ESI-MS-ready IgG preparation from converted serum and the solution was incubated to form an immune complex between the ovalbumin-derived epitope peptide and the anti-ovalbumin antibody. Then, the entire mixture of proteins and peptides was directly electrosprayed. Sorting of ions in the mass spectrometer's gas phase, dissociation of the immune complex ions by collision induced dissociation, and recording of the epitope peptide ion which had been released from the immune complex revealed the presence of the anti-ovalbumin antibody in converted serum. Mass determination of the complex-released epitope peptide ion with isotope resolution is highly accurate, guaranteeing high specificity of this novel seroconversion analysis approach which is termed Intact Transition Epitope Mapping – Serological Inspections by Epitope EXtraction (ITEM – SIX).

INSTRUMENT(S):

ORGANISM(S): Gallus Gallus (chicken)

SUBMITTER: Michael Kreutzer  

LAB HEAD: Prof. Dr. Michael O. Glocker

PROVIDER: PXD040754 | Pride | 2023-03-31

REPOSITORIES: Pride