In-depth method for the characterization of glycosylation in manufactured recombinant monoclonal antibody drugs.
The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. Characterization is important but complicated by the intricate structures, microheterogeneity, and the limitations of current tools for structural analysis. In this study, we developed a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs. A library of over 70 structures was developed for the rapid characterization of rMab. N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. The complete structures were obtained through exoglycosidase sequencing. The results showed that most of the N-glycans between different antibodies are nearly the same with different abundances. The utility of this library enables one to identify structures in a rapid manner by matching LC retention times and accurate masses.
Department of Chemistry, University of California , One Shields Avenue, Davis, California 95616, United States.
Ting
T
Song
Sureyya
S
Ozcan
Alicia
A
Becker
Carlito B
CB
Lebrilla
D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001798:Blood Proteins / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry; D006031:Glycosylation; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry
D000911:Antibodies, Monoclonal; D001798:Blood Proteins; D006023:Glycoproteins; D011994:Recombinant Proteins