Abstract
Macrophages will play an important role in future diagnostics and immunotherapies of cancer. However, this demands to selectively capture and sort different subpopulations, which remains a challenge due to their innate ability to bind to a wide range of interfaces indiscriminately. The main obstacle here is the lack of interfaces combining sufficient antifouling properties with the display of specific binding sites allowing sorting and quantification. Herein, as a proof of principle means, it is introduced to pattern interfaces to locally and selective capture macrophages. The repellent coating is based on antifouling polymer brushes, which can be functionalized. Arrays of binding sites are constructed by microchannel cantilever spotting. Those structures are tested for the isolation of different macrophage subtypes, especially polarized anti‐inflammatory macrophages of the M2 type which can be found associated to tumors (“tumor associated macrophages”; TAMs). Using macrophages as a model system, it is demonstrated that the newly developed surfaces and patterns are efficient for specifically trapping targeted cells and can be useful for further development of therapeutic or diagnostic purposes in the future.
Highlights
Macrophages will play an important role in future diagnostics and immunoways
Our studies demonstrated that the hierarchical antifouling polymer brushes could repel the adhesion of macrophages, one of the most adherent cell types.[25,26]
The polymer brushes were functionalized to introduce specific capture arrays based on biotin–DBCO immobilization with subsequent macrophage capture over a biotin–streptavidin–biotin sandwich
Summary
Hierarchical polymer brushes were synthesized from silicon and glass substrates. The brushes consist of a bottom block (poly(oligoethylene glycol methylether methacrylate)) with repellent properties and a top block which can be functionalized by click chemistry (refer to Figure 1). The synthesis of the polymer brushes can be divided into three steps: 1) formation of SAM of initiator, 2) grafting of the antifouling block, and 3) grafting of the azide-functional block. Undecyl 2-bromo-2-methylpropanoate, was selected because of its ability to form well-organized and stable monolayers on different substrates.[60] The polymer brushes were grafted from the initiator layer using surface-initiated atom transfer radical polymerization (SI-ATRP). This type of polymerization provides living end groups which are crucial for successful grafting of the top block. The dry ellipsometric thickness of the first block was 20 nm, further increasing to 24 nm after grafting of the second block, and remained the same after functionalization of brushes with azide groups (Table S1, Supporting Information)
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