Publication: Research - peer-review › Journal article – Annual report year: 2011
Standard cell culture plastic was surface modified by passive adsorption or covalent attachment of interleukin (IL)-4 and investigated for its ability to induce differentiation of human monocytes into mature dendritic cells, a process dose-dependently regulated by IL-4. Covalent attachment of IL-4 proceeded via anthraquinone photochemistry to introduce amine functionalities at the surface followed by coupling of IL-4 through a bifunctional amine-reactive linker. X-ray photoelectron spectroscopy showed that undesirable multilayer formation of the photoactive compound could be avoided by reaction in water instead of phosphate-buffered saline. Passively adsorbed IL-4 was observed to induce differentiation to dendritic cells, but analysis of cell culture supernatants revealed that leakage of IL-4 into solution could account for the differentiation observed. Covalent attachment resulted in bound IL-4 at similar concentrations to the passive adsorption process, as measured by enzyme-linked immunosorbent assays, and the bound IL-4 did not leak into solution to any measurable extent during cell culture. However, covalently bound IL-4 was incapable of inducing monocyte differentiation. This may be caused by IL-4 denaturation or improper epitope presentation induced by the immobilization process, or by biological irresponsiveness of monocytes to IL-4 in immobilized formats.
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- Immobilization process, Bifunctional, Cells, surface modification, monocyte, Cell culture, Amine functionality, Enzyme linked immunosorbent assay, X ray photoelectron spectroscopy, IL-4, Dendritic cells, Animal cell culture, cell culture plastic, Saline water, Standard cell, Covalent attachment, Phosphate-buffered salines, Supernatants, Body fluids, differentiation, Adsorption process, Multilayer formation, Human monocytes, Photoactive compounds, Ketones, Electric batteries, Interleukin-4, Adsorption, Surface-modified