Inhibition of mannosidase in hybridomas yields monoclonal antibodies with greater capacity for carbohydrate labeling
| Autor: | Cynthia Long, R W Gillette, T J McKearn, R B Simonson, J D Rodwell, M E Ultee |
|---|---|
| Rok vydání: | 1988 |
| Předmět: |
Mannosidase
chemistry.chemical_classification biology medicine.drug_class Biochemistry (medical) Clinical Biochemistry Mannose Monoclonal antibody Molecular biology Immunoglobulin G chemistry.chemical_compound chemistry Biochemistry Antigen medicine biology.protein Antibody Glycoprotein Polyacrylamide gel electrophoresis |
| Zdroj: | Clinical Chemistry. 34:1713-1716 |
| ISSN: | 1530-8561 0009-9147 |
| DOI: | 10.1093/clinchem/34.9.1709 |
| Popis: | Labeling an antibody site specifically through its carbohydrate residues preserves more of its antigen-binding activity than does labeling through protein moieties. To boost the amount of immunoglobulin G carbohydrate capable of being labeled, we treated hybridoma cells with a mannosidase inhibitor, deoxymannojirimycin (dMM). Polyacrylamide gel electrophoresis showed formation of a glycoprotein with high mannose content, in that endo-beta-N-acetylglucosaminidase H (EC 3.2.1.96) could digest the antibody from the dMM-treated cells, but not from control cultures. Carbohydrate analysis confirmed this conclusion, indicating that the antibody from the dMM-treated cells had twice as much mannose as did the control antibody. The glucosamine content of the treated-cells' antibodies was half that of the control, and no additional carbohydrate residues were detectable in the antibodies secreted by the dMM-treated cells. We conjugated both the dMM and control antibodies through their carbohydrate to a chelator. In labeling, the dMM antibody conjugate incorporated approximately threefold as much 111In isotope as the control conjugate. The two labeled antibodies were injected into mice and showed similar organ distributions. |
| Databáze: | OpenAIRE |
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