Targeted delivery of saporin toxin by monoclonal antibody to the transcobalamin receptor, TCblR/CD320.
Cellular uptake of cobalamin (Cbl) occurs by endocytosis of transcobalamin saturated with Cbl by the transcobalamin receptor (TCblR/CD320). The cell cycle-associated overexpression of this receptor in many cancer cells provides a suitable target for delivering chemotherapeutic drugs and cytotoxic molecules to these cells while minimizing the effect on the normal cell population. We have used monoclonal antibodies to the extracellular domain of TCblR to deliver saporin-conjugated secondary antibody to various cell lines propagating in culture. A molar ratio of 2.5:10 nmol/L of primary:secondary antibody concentration was identified as the lowest concentration needed to produce the optimum cytotoxic effect. The effect was more pronounced when cells were seeded at lower density, suggesting lack of cell division in a fraction of the cells at higher density as the likely explanation. Cells in suspension culture, such as K562 and U266 cells, were more severely affected than adherent cultures, such as SW48 and KB cells. This differential effect of the anti-TCblR-saporin antibody conjugate and the ability of an anti-TCblR antibody to target proliferating cells were further evident by the virtual lack of any effect on primary skin fibroblasts and minimal effect on bone marrow cells. These results indicate that preferential targeting of some cancer cells could be accomplished through the TCblR.
Quadros
Edward V
EV
Department of Medicine, SUNY Downstate Medical Center, Brooklyn, New York, USA. Edward.Quadros@downstate.edu
Nakayama
Yasumi
Y
Sequeira
Jeffrey M
JM
D000911:Antibodies, Monoclonal / Q000008:administration & dosage*; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D000922:Immunotoxins / Q000008:administration & dosage; D020014:K562 Cells; D058990:Molecular Targeted Therapy / Q000379:methods*; D009369:Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D054789:Ribosome Inactivating Proteins, Type 1 / Q000008:administration & dosage*; D000076985:Saporins; D014162:Transfection
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; C538894:CD320 protein, human; D000922:Immunotoxins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D054789:Ribosome Inactivating Proteins, Type 1; C028176:transcobalamin receptor; D049452:Green Fluorescent Proteins; D000076985:Saporins