A streptavidin mutant useful for directed immobilization on solid surfaces

被引:42
作者
Reznik, GO [1 ]
Vajda, S
Cantor, CR
Sano, T
机构
[1] Boston Univ, Ctr Adv Biotechnol, Boston, MA 02215 USA
[2] Boston Univ, Dept Biomed Engn, Boston, MA 02215 USA
[3] Boston Univ, Dept Pharmacol & Expt Therapeut, Boston, MA 02215 USA
[4] Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Dept Radiol,Ctr Mol Imaging Diagnosis & Therapy, Boston, MA 02215 USA
[5] Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Dept Radiol,Basic Sci Lab, Boston, MA 02215 USA
关键词
D O I
10.1021/bc015507t
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A streptavidin mutant has been designed and produced that allows the specific, covalent immobilization of streptavidin on solid surfaces. This streptavidin mutant was constructed by fusing a six-residue sequence, containing a single cysteine, to the carboxyl terminus of streptavidin. Because this mutant has no other cysteine residues, the reactive sulfhydryl group of the cysteine residue serves as a unique immobilization site for corrugation using sulfhydryl chemistry. This streptavidin mutant was efficiently immobilized on maleimide-coated solid surfaces via, its unique immobilization site. Characterization of the immobilized streptavidin mutant for the ability to bind to biotinylated macromolecules and the dissociation rates of bound biotin showed that the biotin-binding properties of this mutant were minimally affected by immobilization on solid surfaces. This streptavidin could be readily incorporated into a wide variety of solid-phase diagnostic tests and biomedical assays. This could enhance the performance of streptavidin-based solid-phase assay systems.
引用
收藏
页码:1000 / 1004
页数:5
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