Microheterogeneity of a purified IgG1 due to asymmetric FAB glycosylation

Microheterogeneity of a purified IgG1 due to asymmetric FAB glycosylation

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...

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Veröffentlicht in:Molecular immunology 1992-06, Vol.29 (6), p.751-758
Hauptverfasser: GREBENAU, R. C, GOLDENBERG, D. M, CHIEN-HSING CHANG, KOCH, G. A, GOLD, D. V, KUNZ, A, HANSEN, H. J
Format: Artikel
Sprache:eng
Schlagworte:
Animals
Antibodies, immunoglobulins
Antibodies, Monoclonal - chemistry
Biological and medical sciences
Blotting, Western
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Glycosylation
Immunoglobulin Fab Fragments - chemistry
Immunoglobulin G - chemistry
Immunoglobulin Heavy Chains - chemistry
In Vitro Techniques
Macromolecular Substances
Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase - pharmacology
Mice
Molecular immunology
Structure
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Zusammenfassung: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.
ISSN:0161-5890
1872-9142
DOI:10.1016/0161-5890(92)90185-Z