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Selective immobilization of Sonic hedgehog on benzylguanine terminated patterned self-assembled monolayers

Paper ID Volume ID Publish Year Pages File Format Full-Text
7790 564 2011 10 PDF Available
Title
Selective immobilization of Sonic hedgehog on benzylguanine terminated patterned self-assembled monolayers
Abstract

Patterned two-component, self-assembled monolayers on gold were produced by UV lithography. An oligo(ethylene glycol) terminated disulfide served as inert matrix reducing unspecific protein adsorption and cell adhesion. The second component of the self-assembled monolayer (SAM) presented a benzylguanine moiety for the immobilization of Sonic hedgehog (Shh) fused to a mutant O6-alkylguanine-DNA alkyltransferase (SNAP-tag™). The enzymatic activity of the SNAP-tag allows selective and covalent immobilization of the linked Shh. Time-of-flight secondary ion mass spectrometry verified the correct lateral distribution of the benzylguanine head groups in the patterned SAM. The quantification of unspecific and specific protein binding to mixed SAMs showed increased adsorption of albumin with increasing benzylguanine/(ethylene glycol) ratios. However, the immobilization of SNAP-tagged Shh was not blocked by pre-adsorbed albumin. Furthermore, the obtained micro-patterned substrates permitted direct immobilization of SNAP-tagged Shh even in the presence of many competing proteins from conditioned media of transfected HEK293 cells. Therefore, the presented system is suited for the controlled immobilization of fusion proteins from complex mixtures avoiding purification steps.

Keywords
Site-directed immobilization; Benzylguanine transferase tag; Sonic hedgehog; Micro-patterning; Self-assembled monolayers; Time-of-flight secondary ion mass spectrometry
First Page Preview
Selective immobilization of Sonic hedgehog on benzylguanine terminated patterned self-assembled monolayers
Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 32, Issue 28, October 2011, Pages 6719–6728
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering