Streptavidin Coverage on Biotinylated Surfaces

P. H.Erik Hamming, Jurriaan Huskens*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

7 Citations (Scopus)
74 Downloads (Pure)


Biosensors and other biological platform technologies require the functionalization of their surface with receptors to enhance affinity and selectivity. Control over the functionalization density is required to tune the platform's properties. Streptavidin (SAv) monolayers are widely used to immobilize biotinylated proteins, receptors, and DNA. The SAv density on a surface can be varied easily, but the predictability is dependent on the method by which the SAv is immobilized. In this study we show a method to quantitatively predict the SAv coverage on biotinylated surfaces. The method is validated by measuring the SAv coverage on supported lipid bilayers with a range of biotin contents and two different main phase lipids and by using quartz crystal microbalance and localized surface plasmon resonance. We explore a predictive model of the biotin-dependent SAv coverage without any fit parameters. Model and data allow to predict the SAv coverage based on the biotin coverage, in both the low- and high-density regimes. This is of special importance in applications with multivalent binding where control over surface receptor density is required, but a direct measurement is not possible.

Original languageEnglish
Pages (from-to)58114-58123
Number of pages10
JournalACS Applied Materials and Interfaces
Issue number48
Publication statusPublished - 8 Dec 2021


  • biotin
  • localized surface plasmon resonance (LSPR)
  • quartz crystal microbalance (QCM)
  • self-assembled monolayer (SAM)
  • streptavidin (SAv)
  • supported lipid bilayer (SLB)
  • surface coverage
  • UT-Hybrid-D


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