Boron-enriched streptavidin potentially useful as a component of boron carriers for neutron capture therapy of cancer.
A boron-enriched streptavidin has been prepared by chemical conjugation of a boron-rich compound, B(12)H(11)SH(2)(-) (BSH), to a genetically engineered streptavidin variant. The streptavidin variant used has 20 cysteine residues per molecule, derived from a C-terminal cysteine stretch consisting of five cysteine residues per subunit. Because natural streptavidin has no cysteine residues, the reactive sulfhydryl groups of the cysteine stretch serve as unique conjugation sites for sulfhydryl chemistry. BSH was conjugated irreversibly to the sulfhydryl groups of the streptavidin variant via a sulfhydryl-specific homobifunctional chemical cross-linker. Quantitative boron analysis indicates that the resulting streptavidin-BSH conjugate carries approximately 230 boron atoms/molecule. This indicates that the chemical conjugation of BSH to the streptavidin variant was highly specific and efficient because this method should allow the conjugation of a maximum of 240 boron atoms/streptavidin molecule. This boron-enriched streptavidin retained both full biotin-binding ability and tetrameric structure, suggesting that the conjugation of BSH has little, if any, effect on the fundamental properties of streptavidin. This boron-enriched streptavidin should be very useful as a component of targetable boron carriers for neutron capture therapy of cancer. For example, a monoclonal antibody against a tumor-associated antigen can be attached tightly to the boron-enriched streptavidin upon simple biotinylation, and the resulting conjugate could be used to target boron to tumor cells on which the tumor-associated antigen is overexpressed.
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Center for Molecular Imaging Diagnosis and Therapy, and Basic Science Laboratory, Department of Radiology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
D000818:Animals; D001710:Biotin / Q000378:metabolism; D001894:Borohydrides / Q000008:administration & dosage* / Q000737:chemistry* / Q000378:metabolism; D001895:Boron / Q000008:administration & dosage* / Q000737:chemistry* / Q000378:metabolism; D016754:Boron Neutron Capture Therapy / Q000379:methods*; D002417:Cattle; D002846:Chromatography, Affinity; D003432:Cross-Linking Reagents / Q000737:chemistry; D004337:Drug Carriers; D004591:Electrophoresis, Polyacrylamide Gel; D019809:Streptavidin / Q000031:analogs & derivatives*; D013438:Sulfhydryl Compounds / Q000008:administration & dosage* / Q000737:chemistry* / Q000378:metabolism; D013439:Sulfhydryl Reagents / Q000737:chemistry
D001894:Borohydrides; D003432:Cross-Linking Reagents; D004337:Drug Carriers; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; C014651:mercaptoundecahydrododecaborate; D001710:Biotin; D019809:Streptavidin; D001895:Boron