TY - GEN
T1 - Arrayed nanohydrogels for protein array technology
AU - Saaem, I.
AU - Papasotiropoulos, V.
AU - Wang, T.
AU - Soteropoulos, P.
AU - Libera, M.
PY - 2005
Y1 - 2005
N2 - This paper describes studies of surface-patterned nanohydrogels and their use as possible substrates for high-density and high-sensitivity protein arrays. Nanohydrogels, approximately 200 nm in diameter, were created by locally crosslinking dry amine-terminated poly(ethylene glycol) [PEG] (5000 Da) thin films using a focused electron beam. These gels then had a dry height of 50 nm and a swell ratio of about five. They were patterned into arrays with approximately 1 μm inter-gel spacing. These arrayed gels were functionalized with Zinc Finger 9 (ZNF9), a nucleic-acid binding protein. As part of ongoing research, we are interested in how the performance of the functionalized gels compares to that of current commercial substrates. We show, using an assay that binds α-GST antibody to mediate attachment of GST-tagged ZNF9, that nanohydrogels have a consistently higher combination of fluorescent signal and signal-to-noise (SNR) ratio than the comparable commercial substrates. We further show that the SNR characteristic of our assay degrades slower on the nanohydrogel array than on a comparable microarray printed on an epoxide substrate, and we speculate that the difference is in part attributable to the fact that the nanohydrogel array presents a softer and more hydrophilic surface.
AB - This paper describes studies of surface-patterned nanohydrogels and their use as possible substrates for high-density and high-sensitivity protein arrays. Nanohydrogels, approximately 200 nm in diameter, were created by locally crosslinking dry amine-terminated poly(ethylene glycol) [PEG] (5000 Da) thin films using a focused electron beam. These gels then had a dry height of 50 nm and a swell ratio of about five. They were patterned into arrays with approximately 1 μm inter-gel spacing. These arrayed gels were functionalized with Zinc Finger 9 (ZNF9), a nucleic-acid binding protein. As part of ongoing research, we are interested in how the performance of the functionalized gels compares to that of current commercial substrates. We show, using an assay that binds α-GST antibody to mediate attachment of GST-tagged ZNF9, that nanohydrogels have a consistently higher combination of fluorescent signal and signal-to-noise (SNR) ratio than the comparable commercial substrates. We further show that the SNR characteristic of our assay degrades slower on the nanohydrogel array than on a comparable microarray printed on an epoxide substrate, and we speculate that the difference is in part attributable to the fact that the nanohydrogel array presents a softer and more hydrophilic surface.
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U2 - 10.1557/proc-0897-j06-07
DO - 10.1557/proc-0897-j06-07
M3 - Conference contribution
AN - SCOPUS:34249947287
SN - 1558998527
SN - 9781558998520
T3 - Materials Research Society Symposium Proceedings
SP - 117
EP - 122
BT - Biomimetic Polymers and Gels
T2 - 2005 MRS Fall Meeting
Y2 - 28 January 2005 through 2 December 2005
ER -