A DNA-Mediated Homogeneous Binding Assay for Proteins and Small Molecules

TY - JOUR

T1 - A DNA-Mediated Homogeneous Binding Assay for Proteins and Small Molecules

AU - Zhang, Zhao

AU - Hejesen, Christian

AU - Kjelstrup, Michael Brøndum

AU - Birkedal, Victoria

AU - Gothelf, Kurt Vesterager

PY - 2014/8/6

Y1 - 2014/8/6

N2 - Optical detection of molecular targets typically requires immobilization, separation, or chemical or enzymatic processing. An important exception is aptamers that allow optical detection in solution based on conformational changes. This method, however, requires the laborious selection of aptamers with high target specificity and affinity, and the ability to undergo the required conformational changes. Here we report on an alternative generic scheme for detecting small molecules and proteins in solution based on a shift in the equilibrium of DNA-based strand displacement competition reaction. The shift occurs upon binding of a protein, for example, an antibody to its target. We demonstrate nanomolar detection of small molecules such as biotin, digoxigenin, vitamin D, and folate, in buffer and in plasma. The method is flexible, and we also show nanomolar detection of the respective antibodies or protein targets of these molecules. The detection scheme provides a generic alternative to aptamers for detection of analytes.

AB - Optical detection of molecular targets typically requires immobilization, separation, or chemical or enzymatic processing. An important exception is aptamers that allow optical detection in solution based on conformational changes. This method, however, requires the laborious selection of aptamers with high target specificity and affinity, and the ability to undergo the required conformational changes. Here we report on an alternative generic scheme for detecting small molecules and proteins in solution based on a shift in the equilibrium of DNA-based strand displacement competition reaction. The shift occurs upon binding of a protein, for example, an antibody to its target. We demonstrate nanomolar detection of small molecules such as biotin, digoxigenin, vitamin D, and folate, in buffer and in plasma. The method is flexible, and we also show nanomolar detection of the respective antibodies or protein targets of these molecules. The detection scheme provides a generic alternative to aptamers for detection of analytes.

U2 - 10.1021/ja505519b

DO - 10.1021/ja505519b

M3 - Journal article

C2 - 25051012

SN - 0002-7863

VL - 136

SP - 11115

EP - 11120

JO - Journal of American Chemical Society

JF - Journal of American Chemical Society

IS - 31

ER -