Practical application of bioluminescence enzyme immunoassay using enhancer for firefly luciferin-luciferase bioluminescence.
Practical application of bioluminescence enzyme immunoassay using enhancer for firefly luciferin-luciferase bioluminescence.
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DOI：
10.1002/bio.1200
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发表时间：
2011-05
期刊：
影响因子：
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通讯作者：
T. Minekawa;H. Ohkuma;K. Abe;Hiroaki Maekawa;H. Arakawa
中科院分区：
文献类型：
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作者：
T. Minekawa;H. Ohkuma;K. Abe;Hiroaki Maekawa;H. Arakawa
萤火虫荧光素-荧光素酶生物发光以其高量子产率（41.0 ± 7.4%）而闻名。鉴于这种高量子产率，这种生物发光的应用预计将在临床诊断领域有用。这种生物发光的动力学曲线表现出光发射的瞬时上升（<1秒）和快速衰减（5秒后下降至42％）。在本研究中，我们应用了辅酶A、肌苷5'-三磷酸钠盐、三聚磷酸钠和焦磷酸钾四种增强剂来延长发光时间。当使用这些增强剂时，5 秒后发光仅分别下降至 89%、83%、87% 和 82%。这些材料改变了生物发光的动力学曲线，使发光更适合临床应用。它变得更加合适，因为它们通过将发光测量与试剂分配分开来实现高度灵敏的集成和简化设备。利用这些增强剂，我们开发了一种针对乙型肝炎病毒表面抗原 (HBsAg) 的生物发光酶免疫分析 (BLEIA)，该分析采用萤火虫荧光素酶作为标记酶。我们将 HBsAg BLEIA 方法与传统化学发光酶免疫分析方法获得的结果进行比较，发现具有令人满意的相关性（r=0.984，n=118）。
Firefly luciferin-luciferase bioluminescence is known for its high quantum yield (41.0 ± 7.4%). Given this high quantum yield, application of this bioluminescence is expected to be useful in the field of clinical diagnostics. The kinetic profile of this bioluminescence exhibits an instant rise (<1 s) and a rapid decay in light emission (decreased to 42% after 5 s). In this study, we applied four enhancers including coenzyme A, inosine5'-triphosphate sodium salt, sodium tripolyphosphate and potassium pyrophosphate to prolong light emission. When these enhancers were used, luminescence was only decreased to 89, 83, 87 and 82% after 5 s, respectively. These materials modified the kinetic profile of bioluminescence so that the luminescence is more suitable for clinical application. It becomes more suitable because they enable highly sensitive integration and simplification of a device by separating luminescence measurements from dispensing of reagents. Using these enhancers, we then developed a bioluminescent enzyme immunoassay (BLEIA) for hepatitis B virus surface antigen (HBsAg) that employed firefly luciferase as a labeling enzyme. We compared the results obtained from the HBsAg BLEIA method with the conventional chemiluminescent enzyme immunoassay method, and found a satisfactory correlation (r=0.984, n=118).