Endocytosis and degradation of monoclonal antibodies targeting human B-cell malignancies.
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).
Press
O W
OW
Department of Medicine, University of Washington, Seattle.
Farr
A G
AG
Borroz
K I
KI
Anderson
S K
SK
Martin
P J
PJ
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000915:Antibody Affinity; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D001666:Binding Sites, Antibody; D002051:Burkitt Lymphoma / Q000276:immunology*; D002851:Chromatography, High Pressure Liquid; D004705:Endocytosis; D006801:Humans; D007124:Immunoenzyme Techniques; D008223:Lymphoma / Q000276:immunology*; D008854:Microscopy, Electron; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte