Analytical and Bioanalytical Chemistry

, Volume 403, Issue 2, pp 517–526 | Cite as

A hydrogel-based versatile screening platform for specific biomolecular recognition in a well plate format

  • Meike V. Beer
  • Claudia Rech
  • Sylvia Diederichs
  • Kathrin Hahn
  • Kristina Bruellhoff
  • Martin Möller
  • Lothar EllingEmail author
  • Jürgen GrollEmail author
Original Paper


Precise determination of biomolecular interactions in high throughput crucially depends on a surface coating technique that allows immobilization of a variety of interaction partners in a non-interacting environment. We present a one-step hydrogel coating system based on isocyanate functional six-arm poly(ethylene oxide)-based star polymers for commercially available 96-well microtiter plates that combines a straightforward and robust coating application with versatile bio-functionalization. This system generates resistance to unspecific protein adsorption and cell adhesion, as demonstrated with fluorescently labeled bovine serum albumin and primary human dermal fibroblasts (HDF), and high specificity for the assessment of biomolecular recognition processes when ligands are immobilized on this surface. One particular advantage is the wide range of biomolecules that can be immobilized and convert the per se inert coating into a specifically interacting surface. We here demonstrate the immobilization and quantification of a broad range of biochemically important ligands, such as peptide sequences GRGDS and GRGDSK-biotin, the broadly applicable coupler molecule biocytin, the protein fibronectin, and the carbohydrates N-acetylglucosamine and N-acetyllactosamine. A simplified protocol for an enzyme-linked immunosorbent assay was established for the detection and quantification of ligands on the coating surface. Cell adhesion on the peptide and protein-modified surfaces was assessed using HDF. All coatings were applied using a one-step preparation technique, including bioactivation, which makes the system suitable for high-throughput screening in a format that is compatible with the most routinely used testing systems.


We present a hydrogel coating system for well-plates that can be covalently modified with peptides, sugars or proteins by dip coating. These coatings then allow specific interaction screening of the immobilized ligands with peptides, proteins or cells.


Bioassays Immunoassays/ELISA Interaction screening Biofunctionalization Hydrogel coating 96-Well plate format 



Bovine serum albumin


Extracellular matrix


Enzyme-linked immunosorbent assay


Enzyme-linked lectin assay






Lectin II from Griffonia simplicifolia


Primary human dermal fibroblast


Recombinant lectin produced in Escherichia coli BL21


Isophorone diisocyanate




Isocyanate functionalized six-arm star-shaped prepolymer







The authors acknowledge funding by the Deutsche Forschungsgemeinschaft (DFG) within the Research Training Group 1035 “BioInterface” (M.V.B., C.R., M.M., L.E.) and the SPP 1257 “Intelligent Hydrogels” (M.M., J.G.). The authors thank Sarah Krauthausen (Interactive Materials Research Institute (DWI e.V.), Aachen) for excellent technical assistance.


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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Meike V. Beer
    • 1
  • Claudia Rech
    • 2
  • Sylvia Diederichs
    • 3
  • Kathrin Hahn
    • 1
  • Kristina Bruellhoff
    • 3
  • Martin Möller
    • 3
  • Lothar Elling
    • 2
    Email author
  • Jürgen Groll
    • 1
    Email author
  1. 1.Department of Functional Materials in Medicine and DentistryUniversity of WürzburgWürzburgGermany
  2. 2.Laboratory for Biomaterials, Institute for Biotechnology and Helmholtz Institute for Biomedical EngineeringRWTH Aachen UniversityAachenGermany
  3. 3.Interactive Materials Research Institute (DWI e.V.) and Institute of Technical and Macromolecular ChemistryRWTH Aachen UniversityAachenGermany

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