New Architecture for Reagentless, Protein-Based Electrochemical Biosensors.
New Architecture for Reagentless, Protein-Based Electrochemical Biosensors.
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DOI：
10.1021/jacs.7b05953
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发表时间：
2017-09-06
影响因子：
15
通讯作者：
Plaxco KW
中科院分区：
文献类型：
--
作者：
Kang D;Sun S;Kurnik M;Morales D;Dahlquist FW;Plaxco KW
Here we demonstrate a new class of reagentless, single-step sensors for monitoring protein-protein and protein-peptide interactions that is the electrochemical analog of fluorescence polarization (fluorescence anisotropy), a versatile optical approach widely employed to this same end. Our electrochemical sensors consist of a redox-reporter-modified protein (the “receptor”) site-specifically anchored to an electrode via a short, flexible polypeptide linker. Interaction of the protein with its binding partner alters the efficiency with which the attached reporter approaches the electrode surface, thus changing the observed redox current upon voltammetric interrogation. As proof-of-principle we employed the bacterial chemotaxis protein CheY as our receptor. Interaction with either of CheY’s two binding partners, the P2 domain of the chemotaxis kinase, CheA, or the 16-residue “target region” of the flagellar switch protein, FliM, leads to easily measurable changes in output current that trace Langmuir isotherms within error of those seen in solution. Phosphorylation of the electrode-bound CheY decreases its affinity for CheA-P2 and enhances its affinity for FliM in a manner likewise consistent with its behavior in solution. As expected given the proposed sensor signaling mechanism, the magnitude of the binding-induced signal change depends on the placement of the redox reporter on the protein. Following these preliminary studies with CheY we also developed and characterized additional sensors aimed at the detection of specific antibodies using the relevant protein antigens as the receptor. These exhibit excellent detection limits for their targets without the use of reagents or wash steps. This novel, protein-based electrochemical sensing architecture provides a new and potentially promising approach to quantitative, single-step detection of specific proteins and peptides.
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DOI：
10.1039/c3an01079a
发表时间：
2013-10-07
期刊：
The Analyst
影响因子：
--
作者：
Bonham AJ;Paden NG;Ricci F;Plaxco KW
通讯作者：
Plaxco KW
影响因子：
3.2
作者：
Dyer, Collin M.;Dahlquist, Frederick W.
通讯作者：
Dahlquist, Frederick W.
影响因子：
3.2
作者：
Appleby, JL;Bourret, RB
通讯作者：
Bourret, RB
影响因子：
2.9
作者：
LI, JY;SWANSON, RV;WEIS, RM
通讯作者：
WEIS, RM
影响因子：
3
作者：
Lucero, NE;Escobar, GI;Nielsen, K
通讯作者：
Nielsen, K