Simultaneous monitoring of protein adsorption at the solid-liquid interface from sessile solution droplets by ellipsometry and axismmetric drop shape analysis by profile

Jaap Noordmans, Herbert Wormeester, Henk J. Busscher

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Abstract

In this paper two in situ techniques are combined to simultaneously examine protein adsorption at the solid–liquid interface from sessile solution droplets. With axisymmetric drop shape analysis by profile (ADSA-P) the change in solid–liquid interfacial tension is determined, while ellipsometry is employed to measure the amount of protein adsorbed from the same solution droplet at the solid–liquid interface. Three proteins (human serum albumin (HSA), immunoglobulin G (IgG) and fibrinogen (Fb)) were dissolved to a concentration of 0.05 mg ml−1 in PBS (pH 7) and sessile droplets were placed for 2 h on a 47.8 nm thick gold coating on glass. The gold coated glass was positioned onto a quartz prism with immersion oil. The prism was aligned in a rotating analyser ellipsometer and the optical beam was thus allowed to be reflected at the hydrophobic gold surface. The ADSA-P set-up was built in 90° cross-beamed set-up around the prism. By combining the results for the adsorbed amounts and the interfacial tension changes over the two hour adsorption period, two stages in the adsorption process could be distinguished. In the first stage, the adsorbed amounts increase in correspondence with the interfacial tension changes, indicating that the interfacial tension changes are caused by adsorption, whereas in the second stage interfacial tension changes continue despite the adsorbed amounts being constant. Consequently, the second stage must be associated with conformational changes of the adsorbed proteins. For HSA and Fb, the conformational contribution to the interfacial tension changes (7.8 and 5.3 mJ m−2, respectively) were approximately 2-fold the adsorption contribution, while for IgG both were equal around 3 mJ m−2.
Original languageUndefined
Pages (from-to)227-233
Number of pages7
JournalColloids and surfaces B: Biointerfaces
Volume1999
Issue number15
DOIs
Publication statusPublished - 1999

Keywords

  • Protein adsorption
  • Solid–liquid interface
  • Axisymmetric drop shape analysis
  • Ellipsometry
  • METIS-128668
  • IR-74167

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