Tentacle probes: eliminating false positives without sacrificing sensitivity.
Tentacle probes: eliminating false positives without sacrificing sensitivity.
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DOI：
10.1093/nar/gkm113
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发表时间：
2007
影响因子：
14.9
通讯作者：
Caplan MR
中科院分区：
文献类型：
--
作者：
Satterfield BC;West JA;Caplan MR
大多数增加生物传感器特异性或灵敏度的努力导致在总体准确性方面几乎没有增益的权衡。这是因为生物传感器不能比它所基于的亲和相互作用更准确。因此，我们已经开发了一类基于协同性数学原理的新试剂，以提高亲和相互作用的准确性。触手探针（TP）具有类似于分子信标（MB）的发夹结构，用于增强特异性，但通过添加捕获探针进行修饰，以增加动力学和亲和力。它们产生的动力学速率常数比具有相应茎强度的MB快200倍。观察到浓度非依赖性特异性，在高达1 mM浓度的变体分析物下无假阳性。相比之下，MB具有浓度依赖性，并且在变体分析物高于3.88 μM时出现假阳性。这种无标记试剂的快速动力学可能证明对微流体测定中的提取效率、灵敏度和检测时间是重要的。TP的浓度非依赖性特异性可能在起始浓度和纯度未知的测定中非常有用，如生物恐怖或临床护理点诊断的情况。
The majority of efforts to increase specificity or sensitivity in biosensors result in trade-offs with little to no gain in overall accuracy. This is because a biosensor cannot be more accurate than the affinity interaction it is based on. Accordingly, we have developed a new class of reagents based on mathematical principles of cooperativity to enhance the accuracy of the affinity interaction. Tentacle probes (TPs) have a hairpin structure similar to molecular beacons (MBs) for enhanced specificity, but are modified by the addition of a capture probe for increased kinetics and affinity. They produce kinetic rate constants up to 200-fold faster than MB with corresponding stem strengths. Concentration-independent specificity was observed with no false positives at up to 1 mM concentrations of variant analyte. In contrast, MBs were concentration dependent and experienced false positives above 3.88 μM of variant analyte. The fast kinetics of this label-free reagent may prove important for extraction efficiency, hence sensitivity and detection time, in microfluidic assays. The concentration-independent specificity of TPs may prove extremely useful in assays where starting concentrations and purities are unknown as would be the case in bioterror or clinical point of care diagnostics.
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DOI：
10.1073/pnas.88.16.7276
发表时间：
1991-08-01
影响因子：
11.1
作者：
HOLLAND, PM;ABRAMSON, RD;GELFAND, DH
通讯作者：
GELFAND, DH
影响因子：
4.4
作者：
Khodarev, NN;Park, J;Pelizzari, CA
通讯作者：
Pelizzari, CA
影响因子：
14.9
作者：
Gentalen, E;Chee, M
通讯作者：
Chee, M
影响因子：
15
作者：
Kitov, PI;Bundle, DR
通讯作者：
Bundle, DR
DOI：
10.1016/0019-2791(72)90097-3
发表时间：
1972-01-01
期刊：
IMMUNOCHEMISTRY
影响因子：
--
作者：
CROTHERS, DM;METZGER, H
通讯作者：
METZGER, H