Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 11356-11360 |
| Number of pages | 5 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 100 |
| Issue number | 20 |
| DOIs | |
| State | Published - Sep 30 2003 |
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ASJC Scopus subject areas
- Genetics
- General
Cite this
A force-based protein biochip. / Blank, K.; Mai, T.; Gilbert, I.; Schiffmann, S.; Rankl, J.; Zivin, R.; Tackney, C.; Nicolaus, T.; Spinnler, K.; Oesterhelt, F.; Benoit, M.; Clausen-Schaumann, H.; Gaub, H. E.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, No. 20, 30.09.2003, p. 11356-11360.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A force-based protein biochip
AU - Blank, K.
AU - Mai, T.
AU - Gilbert, I.
AU - Schiffmann, S.
AU - Rankl, J.
AU - Zivin, R.
AU - Tackney, C.
AU - Nicolaus, T.
AU - Spinnler, K.
AU - Oesterhelt, F.
AU - Benoit, M.
AU - Clausen-Schaumann, H.
AU - Gaub, H. E.
PY - 2003/9/30
Y1 - 2003/9/30
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0141705409&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0141705409&partnerID=8YFLogxK
U2 - 10.1073/pnas.1934928100
DO - 10.1073/pnas.1934928100
M3 - Article
C2 - 12975526
AN - SCOPUS:0141705409
VL - 100
SP - 11356
EP - 11360
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 20
ER -