Microheterogeneity of a purified IgG1 due to asymmetric Fab glycosylation

  • Mol Immunol. 1992 Jun;29(6):751-8. doi: 10.1016/0161-5890(92)90185-z.
R C Grebenau  1 ,  D M Goldenberg ,  C H Chang ,  G A Koch ,  D V Gold ,  A Kunz ,  H J Hansen
Affiliations
  • 1. Immunomedics Inc., Newark, NJ 07103.
PMID: 1603094 DOI: 10.1016/0161-5890(92)90185-z
Abstract

A murine monoclonal anti-granulocyte IgG1, IMMU-MN3, was seen to exhibit heterogeneity. On reduced SDS-PAGE, the purified antibody appeared as two heavy-chain bands of unequal intensity, and only one light-chain band. Hydrophobic interaction chromatography (HIC) also resolved two populations of the IMMU-MN3 antibody. Based on Concanavalin A affinity chromatography, enzymatic digestion with Endoglycosidase F and carbohydrate analysis, it was found that the heterogeneity detected by SDS-PAGE and HIC was due to differences in glycosylation. Furthermore, sequential gel analysis (non-reduced/reduced) demonstrated that the upper heavy-chain band was asymmetrically glycosylated.

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