Practical applications of high-affinity, albumin-binding proteins from a group G streptococcal isolate.
Practical applications of high-affinity, albumin-binding proteins from a group G streptococcal isolate.
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来自 G 组链球菌分离株的高亲和力白蛋白结合蛋白的实际应用。
DOI：
10.1007/s00253-005-0097-4
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Boyle,MichaelDP
中科院分区：
文献类型：
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作者：
Coyle,EmilyM;Blazer,LeviL;White,AbbyA;Hess,JenniferL;Boyle,MichaelDP
在细菌表面表达的与哺乳动物血浆蛋白具有高度亲和力的结合蛋白，已被证明对纯化和检测人类和动物血浆或血清中几种重要的生物学分子有价值。在这项研究中，我们从牛源的G群链球菌分离物中分离出一个高亲和力的白蛋白结合分子，并证明分离的蛋白可以被生物素化而不失去结合活性，并且可以用作定量测定人血清白蛋白（HSA）的示踪剂。结合蛋白可以固定化，并使用生物素化的HSA示踪剂作为竞争性ELISA格式的选择性捕获试剂。在该检测格式下，检测50%抑制HSA结合的灵敏度小于1 μg/ml。当附着在细菌表面时，这种结合蛋白可以用来消耗人血浆中的白蛋白，通过表面增强激光解吸电离飞行时间质谱分析。
Binding proteins that have high affinities for mammalian plasma proteins that are expressed on the surface of bacteria have proven valuable for the purification and detection of several biologically important molecules from human and animal plasma or serum. In this study, we have isolated a high affinity albumin-binding molecule from a group G streptococcal isolate of bovine origin and have demonstrated that the isolated protein can be biotinylated without loss of binding activity and can be used as a tracer for quantification of human serum albumin (HSA). The binding protein can be immobilized and used as a selective capture reagent in a competitive ELISA format using a biotinylated HSA tracer. In this assay format, the sensitivity of detection for 50% inhibition of binding of HSA was less than 1 μg/ml. When attached to the bacterial surface, this binding protein can be used to deplete albumin from human plasma, as analyzed by surface-enhanced laser desorption ionization time of flight mass spectrometry.