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

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.

Site‐Specific Surface Functionalization via Microchannel Cantilever Spotting (µCS): Comparison between Azide–Alkyne and Thiol–Alkyne Click Chemistry Reactions

“…Functionalization of surfaces and materials through the covalent attachment of organic monolayers has become the focus of much research in the last decade because of numerous applications in chemical/biological sensing, drug delivery, selective gas separation, cancer detection, etc. In this regard, click chemistry reactions – which were introduced by Kolb et al in 2001 – are excellent candidates due to having unique characteristics such as mild reaction conditions, high efficiency, compatibility with different kinds of solvents specially water, high reaction rate, and easy post‐treatment and are thus used for many applications …”

Writing Behavior of Phospholipids in Polymer Pen Lithography (PPL) for Bioactive Micropatterns

“…This system was previously studied for patterning via photolithography [28] and in DPN-generated [29,30,31] lipid structures and could therefore act as an ideal benchmark for our purposes. In particular, the larger surface area of pattern coverage easily achieved by PPL in a timely manner opens up the route for more controlled, parallel experiments by introducing microfluidic chambers, enabling a chip format for mast cell experiments [32].…”

ZnO/Nanocarbons‐Modified Fibrous Scaffolds for Stem Cell‐Based Osteogenic Differentiation

“…[ 14–17 ] Indeed, chemical cues (metal ions, small molecules, and synthetic extracellular matrix) and physical cues (stiffness, micro‐/nanotopography, and physical adhesion property) can efficiently influence MSCs’ functionalities, including regulation of cell adhesion, proliferation, and differentiation. [ 18–29 ] For instance, forces originated from the cell–material interface could change the cell membrane and subsequently affect the cell cytoskeleton, owing to the physical and mechanical interactions between the cells membrane the intracellular mechanoresponsive elements. [ 20,30–33 ] Continuous immersion in Yes‐associated protein (YAP) activation culture environments could decrease the human mesenchymal stem cells (hMSCs) multiple differentiation potency and promote osteogenesis.…”