Preparation of Fab-Fragments of GD2-Specific Antibodies and Analysis of Their Antitumor Activity In Vitro

• Published in: Bulletin of experimental biology and medicine Vol. 154; no. 5; pp. 658 - 663
• Main Authors: Doronin, I. I., Kholodenko, I. V., Molotkovskaya, I. M., Kholodenko, R. V.
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01.03.2013; Springer; Springer Nature B.V
• Subjects: Animals; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - isolation & purification; Antibodies, Monoclonal - therapeutic use; Apoptosis; Ascitic Fluid - immunology; Biomedical and Life Sciences; Biomedicine; Cell Biology; Cell Line, Tumor; Cell Proliferation; Chromatography; Gangliosides; Gangliosides - immunology; Hybridomas - immunology; Immunoglobulin Fab Fragments - immunology; Internal Medicine; Laboratory Medicine; Lymphoma, T-Cell - immunology; Lymphoma, T-Cell - pathology; Lymphoma, T-Cell - therapy; Mice; Monoclonal antibodies; Non-Hodgkin's lymphomas; Oncology; Papain; Pathology; mice T-lymphoma EL-4; apoptosis; monoclonal GD2-specific antibodies; Fab-fragments
• ISSN: 0007-4888; 1573-8221
• DOI: 10.1007/s10517-013-2024-5

Monoclonal antibodies ME361 specific to ganglioside GD2 were isolated from the conditioned medium of hybridoma HB9326 and mouse ascitic fluid by the method of affinity chromatography; their Fab-fragments were obtained by proteolytic cleavage with papain. Evaluation of Fab-fragment specificity by flow cytometry and dot-blot analysis showed that binding effectiveness of fragments with antigens was close to that for the full-length molecule of antigen. It was shown that Fab-fragments and whole antibodies ME361 dose-dependently inhibit the proliferation of cells of mice T-lymphoma EL-4, and induce apoptosis of these cells 24 h after incubation.