Comparison of the cytotoxic potency of T101 Fab, F(ab')2 and whole IgG immunotoxins.
The in vitro killing of the human CEM cell line was studied by using ricin A-chain immunotoxins constructed with either the whole IgG or the Fab and F(ab')2 fragments of the same T101 (anti-CD5) antibody. In the presence of ammonium chloride as an activator, the "whole" immunotoxin as well as the "fragment" immunotoxins did not show any significant difference in the cell killing efficacy. In contrast, without the activator, the efficacy of the T101 immunotoxin was greatly improved when fragments were used. Indeed, at a saturating dose, a cytoreduction of three orders of magnitude was obtained with the fragment immunotoxins vs less than one order of magnitude for the whole immunotoxin, as assessed in a clonogenic assay. This enhancing effect was related to better cell killing kinetics, because with a similar amount of A-chain molecules bound per cell, T101 fragment immunotoxins achieved a twofold faster protein synthesis inactivation rate than the corresponding whole IgG immunotoxin. No significant difference in activity was shown between monovalent (Fab) and divalent (F(ab')2) forms of fragment immunotoxins. The observation that T101 fragment immunotoxins were more potent than intact immunotoxins was extended to another fragment immunotoxin constructed with an antibody (F111.98) directed against a different epitope of the CD5 Ag. In another model (anti-CD22 1G11 antibody on Raji cells), the fragment immunotoxin did not show any superiority over the IgG immunotoxin which was by itself very potent, strongly suggesting an Ag-dependent phenomenon.
Derocq
J M
JM
Department of Immunology, Sanofi Recherche, Montpellier, France.
Casellas
P
P
Laurent
G
G
Ravel
S
S
Vidal
H
H
Jansen
F
F
D000818:Animals; D000911:Antibodies, Monoclonal / Q000633:toxicity*; D000915:Antibody Affinity; D001666:Binding Sites, Antibody; D002413:Cations, Divalent; D002460:Cell Line; D002999:Clone Cells / Q000187:drug effects; D003601:Cytotoxicity Tests, Immunologic; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D000922:Immunotoxins / Q000633:toxicity*; D007700:Kinetics; D051379:Mice; D011500:Protein Synthesis Inhibitors / Q000633:toxicity; D012276:Ricin / Q000633:toxicity; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D002413:Cations, Divalent; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D000922:Immunotoxins; D011500:Protein Synthesis Inhibitors; D012276:Ricin