Endocytosis and degradation of monoclonal antibodies targeting human B-cell malignancies

• Published in: Cancer research (Chicago, Ill.) Vol. 49; no. 17; pp. 4906 - 4912
• Main Authors: PRESS, O. W, FARR, A. G, BORROZ, K. I, ANDERSON, S. K, MARTIN, P. J
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA American Association for Cancer Research 01.09.1989
• Subjects: Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - metabolism; Antibody Affinity; Antigens, Differentiation, B-Lymphocyte - immunology; Antineoplastic agents; B-Lymphocytes - immunology; Binding Sites, Antibody; Biological and medical sciences; Burkitt Lymphoma - immunology; Chromatography, High Pressure Liquid; Endocytosis; General aspects; Humans; Immunoenzyme Techniques; Lymphoma - immunology; Medical sciences; Microscopy, Electron; Pharmacology. Drug treatments; Tumor Cells, Cultured; Human; Cell culture; Tumor specific antigen; Burkitt lymphoma; Isotype; B-Lymphocyte; Binding site; Monoclonal antibody; In vitro; Infection; Degradation; Endocytosis; Biological fixation; Viral disease; Tumor cell
• ISSN: 0008-5472; 1538-7445

Seven murine monoclonal antibodies (MoAbs) recognizing differentiation antigens present on B-lymphocytes were analyzed in preclinical studies for their potential use for antibody-targeted therapy of B-cell malignancies. MoAbs HD37 (anti-CD19), 1F5 (anti-CD20), HD6 (anti-CD22), MB-1 (anti-CD37), G28-5 (anti-CDw40), 7.2 (anti-class II), and DA4-4 (anti-IgM) were studied for their binding avidities, immunoreactivities, isotypes, endocytosis rates, degradation rates, and number of binding sites on Daudi cells. Lineweaver-Burke analyses of 125I-labeled MoAbs demonstrated immunoreactivities ranging from 59 to 92%. Scatchard analyses of 125I-MoAbs demonstrated that five of the antibodies had binding avidities in excess of 10(9) L/M, whereas MoAbs 1F5 and HD37 had avidities of 3-4 x 10(8) L/M. CD20, CD37, mu, and HLA Class II were found to be highly expressed (200,000-400,000 binding sites/cell) on Daudi cells whereas CD19, CD22, and CDw40 were less densely expressed (80,000-100,000 sites/cell). DA4-4 (mu), HD6 (CD22), and G28-5 (CDw40) were rapidly internalized by cells, HD37 (CD19) and MB-1 (CD37) underwent endocytosis at an intermediate rate, and 7.2 (class II) and 1F5 (CD20) were internalized slowly. Trichloroacetic acid precipitation and high-performance liquid chromatography revealed the following relative rates of 125I-MoAb degradation: DA4-4 (mu) greater than HD6 (CD22) greater than HD37 (CD19) greater than G28-5 (CDw40) greater than MB-1 (CD37) greater than 1F5 (CD20) greater than 7.2 (class II).