The surface of solid glass supports for samples in optical microscopy and for biosensors needs to be protein-resistant. A coating of a poly(ethylene glycol) monomethyl ether (mPEG) on the surface of the glass is one promising method for preventing the nonspecific adsorption of proteins. In this study, we have developed a novel technique for achieving an optimal coverage of a glass surface with mPEG to prevent protein adhesion. A clean glass substrate previously treated with (3-aminopropyl)dimethylethoxysilane (APDMES) was treated sequentially with poly(acrylic acid) and subsequently a primary amine derivative of mPEG in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. The resultant glass surface was demonstrated to be highly protein-resistant, and the adsorption of bovine serum albumin decreased to only a few percentage points of that on a glass surface treated with APDMES alone. Furthermore, to extend the present method, we also prepared a glass substrate on which biotinylated poly(ethylene glycol) was cografted with mPEG, and biotinylated myosin subfragment-1 (biotin-S1) was subsequently immobilized on this substrate by biotin/avidin chemistry. Actin filaments were observed to glide on the biotin-S1-coated glass surface in the presence of ATP, and thus, the method is capable of immobilizing the protein specifically without any loss in its biological function.
机构:
Indian Inst Technol, Dept Text Technol, New Delhi 110016, IndiaIndian Inst Technol, Dept Text Technol, New Delhi 110016, India
Gupta, Bhuvanesh
Grover, Navdeep
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Indian Inst Technol, Dept Text Technol, New Delhi 110016, India
Indian Inst Technol, Ctr Biomed Engn, New Delhi 110016, IndiaIndian Inst Technol, Dept Text Technol, New Delhi 110016, India
Grover, Navdeep
Singh, Harpal
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Indian Inst Technol, Ctr Biomed Engn, New Delhi 110016, IndiaIndian Inst Technol, Dept Text Technol, New Delhi 110016, India