Mass spectrometry-based de novo sequence
of the
mouse monoclonal anti-FLAG-M2 antibody. The variable regions of the
heavy (A) and light chains (B) are shown. The MS-based sequence is
shown alongside the previously published sequence in the crystal structure
of the Fab (PDB ID: 2G60) and germline sequence (IMGT-DomainGapAlign; IGHV1-04/IGHJ2; IGKV1-117/IGKJ1).
Differential residues are highlighted by asterisks (*). Exemplary
MS/MS spectra in support of the assigned sequences are shown below
the alignments with protease, precursor charge state, and fragmentation
type indicated. Peptide sequence and fragment coverage are indicated
on top of the spectra, with b/c ions
indicated in blue/teal and y/z ions
in red/orange. The same coloring is used to annotate peaks in the
spectra, with additional peaks such as intact/charge reduced precursors,
neutral losses, and immonium ions indicated in green. Note that to
prevent overlapping peak labels, only a subset of successfully matched
peaks is annotated.