A streptavidin mutant useful for directed immobilization on solid surfaces.
A streptavidin mutant useful for directed immobilization on solid surfaces.
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DOI：
10.1021/bc015507t
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发表时间：
2001-09
影响因子：
4.7
通讯作者：
G. Reznik;S. Vajda;C. Cantor;T. Sano
中科院分区：
文献类型：
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作者：
G. Reznik;S. Vajda;C. Cantor;T. Sano
已经设计并产生了允许将链霉亲和素特异性共价固定在固体表面上的链霉亲和素突变体。该链霉亲和素突变体通过将含有单个半胱氨酸的六个残基序列融合到链霉亲和素的羧基末端来构建。由于该突变体没有其他半胱氨酸残基，因此半胱氨酸残基的反应性巯基用作使用巯基化学进行缀合的独特固定位点。该链霉亲和素突变体通过其独特的固定位点有效地固定在马来酰亚胺包被的固体表面上。固定化链霉亲和素突变体的生物素化大分子结合的能力和结合的生物素的解离速率的表征表明，该突变体的生物素结合特性的影响最小的固定在固体表面上。这种链霉亲和素可以很容易地纳入各种固相诊断测试和生物医学测定。这可以增强基于链霉亲和素的固相测定系统的性能。
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 conjugation 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.