title: Synthesis and characterization of oligomeric nido-carboranyl phosphate diester conjugates to antibody and antibody fragments for potential use in boron neutron capture therapy of solid tumors.
abstract: Antibodies conjugated to oligomeric carboranyl compounds have a high potential as target species for boron neutron capture therapy (BNCT) of solid tumors. As a first step toward developing conjugates with BNCT capabilities, an oligomeric nido-carboranyl phosphate diester (Kane, R. R., Dreschel, K., and Hawthorne, M.F. (1993) J. Am. Chem. Soc. 115, 8853-8854), CB10 (10 nido-carboranes containing 90 boron atoms) with a pseudo-5'-terminal amino group, was conjugated to the anticarcinoembryonic antigen antibody T84.66 and its F(ab') fragment. The homobifunctional linker disuccinimidyl suberate (DSS) was coupled to CB10 via its 5'-terminal amino group followed by removal of excess linker with organic solvent extraction and conjugation with intact antibody. Similarly, the heterobifunctional linker, m-maleimidobenzoyl-N-hydroxysuccinimide (MBS), was coupled to CB10 and conjugated to the hinge region sulfhydryl of the F(ab') fragment of T84.66. The extent of reaction was monitored by the mobility shift of CB10-antibody conjugate on native polyacrylamide gels and the increased susceptibility of the CB10-antibody conjugate to staining with silver nitrate. CB10 was also labeled with radioiodine (131I) in a solid phase reaction with iodogen and used in double-label studies with 125I-labeled antibody. Although free CB10 bound very tightly to gel filtration media such as Sephadex G-25, the CB10-antibody conjugate passed through freely. After separation of CB10-antibody conjugate from free CB10 on Sephadex G-25, molar incorporations of CB10 were calculated. At a molar ratio of 10:1 (CB10:T84.66), greater than 90% of T84.66 and 30% of its F(ab)' fragment were conjugated to CB10.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Division of Immunology, Beckman Research Institute of the City of Hope, Duarte, California 91010.
authors.forename: C J
authors.initials: CJ
authors.lastname: Chen
authors.forename: R R
authors.initials: RR
authors.lastname: Kane
authors.forename: F J
authors.initials: FJ
authors.lastname: Primus
authors.forename: G
authors.initials: G
authors.lastname: Szalai
authors.forename: M F
authors.initials: MF
authors.lastname: Hawthorne
authors.forename: J E
authors.initials: JE
authors.lastname: Shively
mesh_terms: D000818:Animals; D001895:Boron; D001896:Boron Compounds / Q000138:chemical synthesis* / Q000493:pharmacokinetics / Q000627:therapeutic use; D016754:Boron Neutron Capture Therapy; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000532:radiotherapy*; D016499:Radioimmunotherapy; D013388:Succinimides / Q000737:chemistry; D013942:Thymine Nucleotides / Q000138:chemical synthesis* / Q000493:pharmacokinetics / Q000627:therapeutic use; D014018:Tissue Distribution
chemical_list: D001896:Boron Compounds; D002272:Carcinoembryonic Antigen; D007140:Immunoglobulin Fab Fragments; D013388:Succinimides; D013942:Thymine Nucleotides; C092359:carboranyl oligophosphate CB10; C013097:3-maleimidobenzoyl N-hydroxysuccinimide; D001895:Boron; C019358:disuccinimidyl suberate