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Merck

Photochemical immobilization of proteins on microwave-synthesized photoreactive polymers.

Analytical biochemistry (2004-03-23)
Azmi Naqvi, Pradip Nahar
RESUMO

We report a rapid and versatile procedure for the preparation of photoreactive polymers and light-induced immobilization of proteins onto such polymers. Photoreactive controlled-pore glass, silica gel, glass slide, and polystyrene microtiter plate are prepared in 40-60s by microwave irradiation of the respective amino polymers and 1-fluoro-2-nitro-4-azidobenzene. Azido group, now part of the polymer, yields highly reactive nitrene under ultraviolet (UV) light at 365 nm. Thus, when photoreactive polymer and horseradish peroxidase or glucose oxidase are exposed to UV light, the reactive nitrene immobilizes the protein molecules in 10 to 20 min through covalent bonding. As nitrene has a property of inserting into C-H bond, the method may find potential applications for immobilization of biomolecules irrespective of their functional groups.

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Sigma-Aldrich
Azidobenzene solution, ~0.5 M in tert-butyl methyl ether
Sigma-Aldrich
Azidobenzene solution, ~0.5 M in 2-methyltetrahydrofuran, ≥95.0% (HPLC)