Investigation of Reagent Delivery Formats in a Multivalent Malaria Sandwich Immunoassay and Implications for Assay Performance.
Investigation of Reagent Delivery Formats in a Multivalent Malaria Sandwich Immunoassay and Implications for Assay Performance.
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DOI：
10.1021/acs.analchem.5b04222
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发表时间：
2016-02-16
影响因子：
7.4
通讯作者：
Fu E
中科院分区：
文献类型：
--
作者：
Liang T;Robinson R;Houghtaling J;Fridley G;Ramsey SA;Fu E
传统的侧向流测试（LFT），在低资源的护理点（POC）设置中使用的当前标准生物测定形式，具有限制，这阻碍了它们在需要高灵敏度和低检测限的低浓度分析物的测试中的应用。LFT使用预混形式，将预混样品和标记抗体快速一步递送至检测区域。我们比较了两种类型的试剂输送格式的模型系统的夹心免疫分析疟疾蛋白检测的信号特征。预混形式在检测区域内产生均匀的结合特征。相比之下，将样品和标记抗体以顺序形式递送到检测区的解耦产生了不均匀的结合谱，其中大部分信号定位于检测区的上游边缘。以连续和预混形式表征测定响应。根据测定偶联物浓度，连续形式的检测限比预混形式低4 - 10倍。该测定的数学模型定量再现了一组速率常数的实验结合曲线，所述速率常数与实验多价疟疾系统的表面等离子体共振测量和吸光度测量一致。
Conventional lateral flow tests (LFTs), the current standard bioassay format used in low-resource point-of-care (POC) settings, have limitations that have held back their application in the testing of low concentration analytes requiring high sensitivity and low limits of detection. LFTs use a premix format for a rapid one-step delivery of premixed sample and labeled antibody to the detection region. We have compared the signal characteristics of two types of reagent delivery formats in a model system of a sandwich immunoassay for malarial protein detection. The premix format produced a uniform binding profile within the detection region. In contrast, decoupling the delivery of sample and labeled antibody to the detection region in a sequential format produced a non-uniform binding profile in which the majority of the signal was localized to the upstream edge of the detection region. The assay response was characterized in both the sequential and premix formats. The sequential format had a four- to ten-fold lower limit of detection than the premix format, depending on assay conjugate concentration. A mathematical model of the assay quantitatively reproduced the experimental binding profiles for a set of rate constants that were consistent with surface plasmon resonance measurements and absorbance measurements of the experimental multivalent malaria system.
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DOI：
10.1006/meth.2000.1036
发表时间：
2000-09-01
期刊：
METHODS-A COMPANION TO METHODS IN ENZYMOLOGY
影响因子：
--
作者：
Paek, SH;Lee, SH;Kim, YS
通讯作者：
Kim, YS
DOI：
10.3390/s90806084
发表时间：
2009
期刊：
Sensors (Basel, Switzerland)
影响因子：
--
作者：
Assadollahi S;Reininger C;Palkovits R;Pointl P;Schalkhammer T
通讯作者：
Schalkhammer T
影响因子：
2.9
作者：
Qian, SZ;Bau, HH
通讯作者：
Bau, HH
影响因子：
4.3
作者：
Holstein, Carly A.;Chevalier, Aaron;Yager, Paul
通讯作者：
Yager, Paul
DOI：
10.1088/0960-1317/23/6/067003
发表时间：
2013-06-01
影响因子：
2.3
作者：
Spicar-Mihalic, P.;Toley, B.;Fu, E.
通讯作者：
Fu, E.