Reversed-phase high-performance liquid chromatographic tryptic digest peptide map comparisons of monoclonal...

An automatic computer technique was used to compare the retention times and ultraviolet spectra of sixty-two peaks in peptide maps of three monoclonal antibodies against human tumor necrosis factor (TNF) and one monoclonal antibody against recombinant factor VIII. The anti-TNF monoclonal, B6, which has an overlapping epitope with the anti-TNF monoclonal, A10G10, had a 90% peak match with A10G10. The anti-TNF monoclonal, A6, with a different epitope to TNF than A10G10, had only a 60% peak match. The A6 match to A10G10 was similar to the 50% peak match of the anti-factor VIII monoclonal with A10G10. The results of this study suggest that peptide mapping can be used as a quantitative characterization technique for comparing monoclonal antibodies.