A monoclonal antibody against human beta-glucuronidase for application in antibody-directed enzyme prodrug therapy.
The selectivity of anticancer agents may be improved by antibody-directed enzyme prodrug therapy (ADEPT). The immunogenicity of antibody-enzyme conjugates and the low tumor to normal tissue ratio calls for the use of a human enzyme and the development of a monoclonal antibody (MAb) against that enzyme for rapid clearance of the conjugate from the circulation. We isolated beta-glucuronidase from human liver. BALB/c mice were immunized with the roughly purified human liver beta-glucuronidase and we obtained an MAb designated 105. Immunoblotting showed reactivity with native tetrameric human beta-glucuronidase. MAb 105 neither bound to enzyme from bovine liver, rat liver, or mouse liver nor reacted with other human lysosomal enzymes. The antibody appeared to be useful to further purify human beta-glucuronidase from human liver or human placenta to homogeneity by affinity chromatography. MAb 105 did not inhibit the activity of human beta-glucuronidase. When human beta-glucuronidase was injected i.v. into BALB/c mice, the newly generated MAb 105 could indeed accelerate the clearance of the enzyme with a 50% drop in its activity within 5 min.
Department of Medical Oncology, Free University Hospital, Amsterdam, The Netherlands.
H J
HJ
Haisma
M
M
Van Muijen
G
G
Scheffer
R J
RJ
Scheper
H M
HM
Pinedo
E
E
Boven
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000494:pharmacology; D000918:Antibody Specificity; D004789:Enzyme Activation / Q000276:immunology; D005966:Glucuronidase / Q000276:immunology* / Q000302:isolation & purification / Q000493:pharmacokinetics; D006801:Humans; D018796:Immunoconjugates / Q000627:therapeutic use*; D008099:Liver / Q000201:enzymology / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011355:Prodrugs / Q000627:therapeutic use*
D000911:Antibodies, Monoclonal; D018796:Immunoconjugates; D011355:Prodrugs; D005966:Glucuronidase