Full Antibody Sequencing
Proteome Factory is specialized in providing full services for complete antibody and CDR sequencing and characterization using a combination of N-terminal and internal Edman protein sequencing, capillary LC with microfraction collector, MS and MSMS mass spectrometry and de novo peptide sequencing. LC and HC fragments of purified antibody are separately digested by several cleavage methods into peptides to get overlapping amino acid peptide sequences of the protein. The peptide mixtures are analyzed directly by nanoLC-ESI-MSMS and after capLC separation and fractionation of peptides by MALDI-MS / MSMS, nanoESI-MSMS and/or Edman protein sequencing. MSMS peptide fragmentation data are evaluated by de novo peptide sequencing and/or protein database search using Proteome Factory's proprietary Full Antibody Sequencing Software and Database. The full protein amino acid sequence is derived by putting together the peptide puzzle of overlapping peptide sequences.
Optionally validation of protein sequence is performed by SRM (selected reaction monitoring) analyses of peptide sequences which have to be validated by another analytical method.