Click-Chemistry Based Allergen Arrays Generated by Polymer Pen Lithography for Mast Cell Activation Studies

Small. 2016 Oct;12(38):5330-5338. doi: 10.1002/smll.201601623. Epub 2016 Aug 11.

Abstract

The profiling of allergic responses is a powerful tool in biomedical research and in judging therapeutic outcome in patients suffering from allergy. Novel insights into the signaling cascades and easier readouts can be achieved by shifting activation studies of bulk immune cells to the single cell level on patterned surfaces. The functionality of dinitrophenol (DNP) as a hapten in the induction of allergic reactions has allowed the activation process of single mast cells seeded on patterned surfaces to be studied following treatment with allergen specific Immunoglobulin E antibodies. Here, a click-chemistry approach is applied in combination with polymer pen lithography (PPL) to pattern DNP-azide on alkyne-terminated surfaces to generate arrays of allergen. The large area functionalization offered by PPL allows an easy incorporation of such arrays into microfluidic chips. In such a setup, easy handling of cell suspension, incubation process, and read-out by fluorescence microscopy will allow immune cell activation screening to be easily adapted for diagnostics and biomedical research.

Keywords: allergen arrays; mast cell activation; microarrays; microfluidics; polymer pen lithography.

MeSH terms

  • Allergens / chemistry*
  • Animals
  • Cell Count
  • Cell Line
  • Click Chemistry / methods*
  • Immunoglobulin E / metabolism
  • Mast Cells / metabolism*
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Phosphotyrosine / metabolism
  • Polymers / chemistry*
  • Printing / methods*
  • Rats
  • Receptors, Fc / metabolism
  • Rhodamines / chemistry

Substances

  • 5-carboxytetramethylrhodamine succinimidyl ester
  • Allergens
  • Polymers
  • Receptors, Fc
  • Rhodamines
  • Phosphotyrosine
  • Immunoglobulin E