A force-based protein biochip.
A parallel assay for the quantification of single-molecule binding forces was developed based on differential unbinding force measurements where ligand-receptor interactions are compared with the unzipping forces of DNA hybrids. Using the DNA zippers as molecular force sensors, the efficient discrimination between specific and nonspecific interactions was demonstrated for small molecules binding to specific receptors, as well as for protein-protein interactions on protein arrays. Finally, an antibody sandwich assay with different capture antibodies on one chip surface and with the detection antibodies linked to a congruent surface via the DNA zippers was used to capture and quantify a recombinant hepatitis C antigen from solution. In this case, the DNA zippers enable not only discrimination between specific and nonspecific binding, but also allow for the local application of detection antibodies, thereby eliminating false-positive results caused by cross-reactive antibodies and nonspecific binding.
Blank
K
K
nanotype, Lochhamer Schlag 12, 82166 Gräfelfing, Germany.
Mai
T
T
Gilbert
I
I
Schiffmann
S
S
Rankl
J
J
Zivin
R
R
Tackney
C
C
Nicolaus
T
T
Spinnler
K
K
Oesterhelt
F
F
Benoit
M
M
Clausen-Schaumann
H
H
Gaub
H E
HE
D001483:Base Sequence; D015374:Biosensing Techniques; D017931:DNA Primers; D011506:Proteins / Q000737:chemistry*
D017931:DNA Primers; D011506:Proteins