Complete and rapid peptide and glycopeptide mapping of mouse monoclonal antibody by LC/MS/MS using ion trap mass spectrometry.
Complete and rapid peptide and glycopeptide mapping of a mouse monoclonal immunoglobulin (IgG2b) were carried out by liquid chromatography/electrospray ionization ion trap-mass spectrometry/mass spectrometry (LC/ ESI IT-MS/MS). It was possible to obtain spectra of a minor glycopeptide at a quantity as low as 1.8 pmol. Reduced and carboxymethylated mouse antidansyl monoclonal IgG2b (RCM-IgG2b) was digested with lysyl-endopeptidase. Proteolytic peptides were subjected to capillary HPLC separation followed by analysis with an ion trap mass spectrometer. The complete amino acid sequence of the IgG2b was characterized by using LC/ ESI IT-MS/MS. The structures of 12 different types of O-linked oligosaccharides attached to Thr-221AH in the hinge region and those of three major types of N-linked oligosaccharides attached to Asn-297H have been characterized.
Central Research Laboratories, Ajinomoto Company, Inc., Kawasaki, Japan.
K
K
Hirayama
R
R
Yuji
N
N
Yamada
K
K
Kato
Y
Y
Arata
I
I
Shimada
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002853:Chromatography, Liquid / Q000379:methods; D006020:Glycopeptides / Q000032:analysis*; D007074:Immunoglobulin G / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D008969:Molecular Sequence Data; D016853:Muromonab-CD3 / Q000737:chemistry; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000032:analysis*; D012697:Serine Endopeptidases
D000911:Antibodies, Monoclonal; D006020:Glycopeptides; D007074:Immunoglobulin G; D016853:Muromonab-CD3; D010455:Peptides; D012697:Serine Endopeptidases; C036813:lysyl endopeptidase