共查询到20条相似文献,搜索用时 180 毫秒
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目的 构建可在体外应用荧光共振能量转移(FRET)技术检测具有酶活性的前列腺特异抗原(PSA)的生物检测器。方法 构建原核细胞表达质粒,从而在大肠杆菌中表达在供体和受体荧光蛋白之间以PSA特异性识别氨基酸序列SSYYSG连接的融合蛋白。将纯化的融合蛋白依次用糜蛋白酶、胰蛋白酶、商品化的纯品PSA及精液切割,检测其荧光共振能量转移现象的变化。结果 糜蛋白酶、商品化的纯品PSA及精液切割均可有效抑制融合蛋白的FRET现象,并表现出对时间和剂量的依赖关系。而胰蛋白酶在90min内未见FRET抑制现象。结论 本研究构建的FRET检测器可成功检测具有酶活性的PSA ,为活性PSA的检测提供一种可选择的途径。 相似文献
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分子信标在活细胞研究中的应用新进展 总被引:1,自引:0,他引:1
了解分子在空间和时间的相互作用是理解程的关键.分子信标是根据荧光共振能量转移原理设计的一种高灵敏性、高特异性检测工具.分子信标在疾病的诊断及治疗、病原体基因组诊断、药物的研制、筛选方面有着广阔的发展前景.本文综述了分子信标的作用原理、作用机制以及在活细胞研究方面的最新进展. 相似文献
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目的%20构建STAT3二聚化抑制剂筛选模型,为STAT3抑制剂筛选提供实验方法。方法%20分别构建pECFP-N1-STAT3和pEYFP-N1-STAT3的荧光报告载体,利用脂质体转染技术将二者共转入HEK-293T细胞,利用ECFP和EYFP两种荧光蛋白之间能量共振转移,检测磷酸化STAT3分子的二聚化水平,并检测Stattic对二聚化的影响。结果%20成功构建了pECFP-N1-STAT3和pEYFP-N1-STAT3的荧光报告载体。将两种荧光报告载体共转染HEK-293T细胞后,Western%20Blot%20检测结果显示STAT3以及p-STAT3的表达水平明显增加。以458nm波长激发ECFP,其发射波长可激发EYFP。加入STAT3二聚化抑制剂Stattic后,荧光强度降低,且呈现一定的剂量依赖性。结论%20基于荧光共振能量转移技术的STAT3二聚化抑制剂筛选模型构建成功。 相似文献
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作者简述荧光测定法的基本方法与影响因素,着重讨论四个方面的应用.酶免疫分析应用酶免疫分析技术进行药物和违禁药品(柯卡因、海洛因、吗啡等)分析已日趋广泛.在酶免疫分析中,药物的特异性抗体与酶结合形成非活性的酶-抗体-抗原复合物.用放射分析法或荧光法可测定酶活力,后一技术近来已被普遍采用.能量转移技术kosha几十年前指出,如果分子中具有相互重迭的二个产生荧光的部分,分子中的能量 相似文献
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Strachan CJ Rades T Gordon KC Rantanen J 《The Journal of pharmacy and pharmacology》2007,59(2):179-192
Raman spectroscopy is experiencing a surge in interest in solid-state pharmaceutical applications. It is rapid, non-destructive, no sample preparation is required and measurements can be made in aqueous environments. It can be used for not only qualitative, but also quantitative, analysis. In this paper, the use of Raman spectroscopy for quantitative analysis of pharmaceutical solids is reviewed. The technique has been used for chemical and physical form analysis. Physical form analysis has involved quantification of polymorphism, hydrates, the amorphous form and, recently, protein conformation. Initially, simple powder systems were quantified, although this has since extended to complex pharmaceutical formulations, including tablets, capsules, microspheres and suspensions. Formulations have also been analysed through packaging. The characteristics of the technique make it ideal for process monitoring and it has been used to quantify changes in-situ during processes such as wet granulation and batch crystallisation. The theoretical basis of quantitative Raman spectroscopy, common data analysis approaches, including multivariate analysis, and sources of error in quantitative analysis are also discussed. 相似文献
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Recent advances in the application of transmission Raman spectroscopy to pharmaceutical analysis 总被引:1,自引:0,他引:1
This article reviews recent advances in transmission Raman spectroscopy and its applications, from the perspective of pharmaceutical analysis. The emerging concepts enable rapid non-invasive volumetric analysis of pharmaceutical formulations and could lead to many important applications in pharmaceutical settings, including quantitative bulk analysis of intact pharmaceutical tablets and capsules in quality and process control. 相似文献
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以光纤传感常用的两大方法(紫外-可见吸收法和荧光法)为基础,介绍其原理,探讨与其联用技术的特点和在药物分析中国内外的新应用。综述了近年来国内外光纤传感联用溶出装置、流动注射、阀上实验室、芯片技术的特点及应用,为药物分析者提供实际工作的需求,展望了光纤传感在药物分析中,未来发展1+n的联用模式。 相似文献
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超高效液相色谱在药物分析中的应用 总被引:5,自引:0,他引:5
超高效液相色谱(UPLC)是近年来发展的以超高速度、超高分离度、超高灵敏度为特点的新型液相色谱技术。其应用于复杂体系快速分析所表现出的优势,已引起了药物分析工作者的重视。本文对近年来超高效液相色谱在药物分析中的应用进行了综述。 相似文献
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Förster resonance energy transfer (FRET) has been widely used in biological and biomedical research because it can determine molecule or particle interactions within a range of 1–10 nm. The sensitivity and efficiency of FRET strongly depend on the distance between the FRET donor and acceptor. Historically, FRET assays have been used to quantitatively deduce molecular distances. However, another major potential application of the FRET assay has not been fully exploited, that is, the use of FRET signals to quantitatively describe molecular interactive events. In this review, we discuss the use of quantitative FRET assays for the determination of biochemical parameters, such as the protein interaction dissociation constant (Kd), enzymatic velocity (kcat) and Km. We also describe fluorescent microscopy-based quantitative FRET assays for protein interaction affinity determination in cells as well as fluorimeter-based quantitative FRET assays for protein interaction and enzymatic parameter determination in solution. 相似文献
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Ríos A De Castro MD Valcárcel M 《Journal of pharmaceutical and biomedical analysis》1985,3(2):105-112
A review is presented in which the fundamental principles, instrumentation and practical aspects of flow injection analysis (FIA) are discussed. A particular focus is put on applications in pharmaceutical chemistry, which offers a wide scope to the new technique. 相似文献