TY - JOUR
T1 - Immunological identification of fibrinogen in dual-component protein films by AFM imaging
AU - Soman, Pranav
AU - Rice, Zachary
AU - Siedlecki, Christopher A.
N1 - Funding Information:
Financial support for this work was provided by National Heart, Lung, and Blood Institute Contract N01HV48191, the Penn State University BBSI program through a grant from the National Institute of Health and the National Science Foundation (Funding #: NSF EEC-0234026), the Dorothy Foehr Huck and J. Lloyd Huck Institutes of the Life Sciences, and by a grant from the Pennsylvania Department of Health. The Pennsylvania Department of Health specifically disclaims responsibility for any analyses, interpretations or conclusions. The authors thank Dr. Keith Milner for preparing the silicone stamps for micro-contact printing and Dr. John Flanagan for assistance with HPLC purification. The authors would also like to thank Dr. Li-Chong Xu and Dr. Aashiish Agnihotri for valuable technical discussions.
PY - 2008/10
Y1 - 2008/10
N2 - The success of long-term blood-contacting implanted devices is largely dependent upon the interaction of the blood components with the device biomaterial surface. The ability to study these interactions has been hindered by a lack of methods to measure single-molecule interactions in complex multi-protein environments similar to the environment found in vivo. In this paper, we demonstrate the use of atomic force microscopy (AFM) in conjunction with gold nanolabels to detect the protein fibrinogen under aqueous conditions without the topographical clues usually necessary for high resolution visualization. BSA was patterned onto both muscovite mica and plasma-treated polydimethylsiloxane (PDMS) substrates and these test substrates were subsequently backfilled with fibrinogen to yield a featureless protein layer. The fibrinogen in this dual-protein layer was detected using high resolution AFM imaging following infusion of anti-fibrinogen conjugated with nanogold particles. This AFM immuno-detection technique will potentially be applicable to complex multi-component protein films adsorbed on clinically relevant polymers used in medical devices.
AB - The success of long-term blood-contacting implanted devices is largely dependent upon the interaction of the blood components with the device biomaterial surface. The ability to study these interactions has been hindered by a lack of methods to measure single-molecule interactions in complex multi-protein environments similar to the environment found in vivo. In this paper, we demonstrate the use of atomic force microscopy (AFM) in conjunction with gold nanolabels to detect the protein fibrinogen under aqueous conditions without the topographical clues usually necessary for high resolution visualization. BSA was patterned onto both muscovite mica and plasma-treated polydimethylsiloxane (PDMS) substrates and these test substrates were subsequently backfilled with fibrinogen to yield a featureless protein layer. The fibrinogen in this dual-protein layer was detected using high resolution AFM imaging following infusion of anti-fibrinogen conjugated with nanogold particles. This AFM immuno-detection technique will potentially be applicable to complex multi-component protein films adsorbed on clinically relevant polymers used in medical devices.
UR - http://www.scopus.com/inward/record.url?scp=48749103309&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=48749103309&partnerID=8YFLogxK
U2 - 10.1016/j.micron.2007.12.013
DO - 10.1016/j.micron.2007.12.013
M3 - Article
C2 - 18294855
AN - SCOPUS:48749103309
SN - 0968-4328
VL - 39
SP - 832
EP - 842
JO - Zeitschrift fur wissenschaftliche Mikroskopie und mikroskopische Technik
JF - Zeitschrift fur wissenschaftliche Mikroskopie und mikroskopische Technik
IS - 7
ER -