Landing Rate Measurements to Detect Fibrinogen Adsorption to Non-fouling Surfaces
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Rapid advances in non-fouling surface technology have pushed the performance of novel coatings toward the detection limit of established protein density quantification techniques. Hence, there is an urgent need for more sensitive detection strategies. Previously we demonstrated that landing rate measurements of microtubules can reveal kinesin surface coverages between 0.1 and 10 μm−2. In this report, we quantify the binding kinetics of highly fluorescent markers to surface-adhered proteins and demonstrate the of protein surface densities in the range of 0.1–1000 μm−2. We utilize this technique to measure kinesin densities on casein-coated glass surfaces and fibrinogen densities on non-fouling polyethylene glycol methacrylate (PEGMA) surfaces. The use of nanospheres (i) potentially permits the detection of a variety of adsorbed proteins, (ii) facilitates the determination of the landing rate due to their uniformity, and (iii) extends the dynamic range of the method due to their small size.
KeywordsBiomaterials Coatings Fibrinogen Protein adsorption
The authors thank Amit Singh of PERC, University of Florida, and Siheng He for helpful discussions. H.H. was supported by NSF Award DMR 1015486.
Conflict of interest
The authors do not have any conflict of interest to declare.
- 1.Adamczyk, Z., J. Barbasz, and M. Ciesla. Kinetics of fibrinogen adsorption on hydrophilic substrates. Langmuir 27:6868–6878, 2011.Google Scholar
- 2.Adamczyk, Z., P. Belouschek, and D. Lorenz. Electrostatic interactions of bodies bearing thin double-layers, I. General formulation. Ber. Bunsenges. Phys. Chem. 94:1483–1492, 1990.Google Scholar
- 5.Black, J. Biological Performance of Materials (4th ed.). Boca Raton: Taylor & Francis, 2006.Google Scholar
- 6.Cao, L., M. Chang, C.-Y. Lee, D. G. Castner, S. Sukavaneshvar, B. D. Ratner, and T. A. Horbett. Plasma-deposited tetraglyme surfaces greatly reduce total blood protein adsorption, contact activation, platelet adhesion, platelet procoagulant activity, and in vitro thrombus deposition. J. Biomed. Mater. Res. A 81A:827–837, 2007.CrossRefGoogle Scholar
- 10.Estephan, Z. G., J. B. Schlenoff, and P. S. Schlenoff. Zwitteration as an alternative to PEGylation. Langmuir 27:6794–6800, 2011.Google Scholar
- 25.Mosesson, M. W. Fibrin polymerization and its regulatory role in hemostasis. J. Lab. Clin. Med. 116:8–17, 1990.Google Scholar