Patterning Functional Proteins with High Selectivity for Biosensor Applications

被引：0
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作者：
Li, Nan [1 ]
Ho, Chih-Ming [1 ]
机构：
[1] University of California, Los Angeles, CA, United States
来源：
JALA - Journal of the Association for Laboratory Automation | 2008年 / 13卷 / 04期
关键词：
In this article; two-dimensional hexamethyldisilazane (HMDS) micropatterns were generated on glass substrates using photolithographic techniques for the assembly of functional proteins. The non-HMDS patterned areas were backfilled with poly(ethylene glycol) (PEG) silane to reduce the nonspecific protein adsorption. The hydrophobic methyl-terminated HMDS monolayer was verified to be favorable for physical protein adsorption with bovine serum albumin (BSA). The PEG-silane derivatized surface significantly reduced the BSA nonspecific binding by 97% compared to the pristine glass substrate so that high patterning selectivity was achieved. A universal streptavidin template was generated using preadsorbed biotinylated BSA on HMDS surface to sequentially bind additional biotinylated antibodies. Using this patterning strategy; the biotinylated goat anti-mouse (biotin-GAM) antibodies can be specifically recognized by the fluorescently labeled mouse immunoglobulin G; which indicated that the immobilized biotin-GAM was still bioactive. Also; the immobilized alkaline phosphatase was demonstrated to retain its enzymatic functionality by the ability to convert its fluorogenic substrate fluorescein diphosphate into fluorescent products. This simple and effective protein patterning technique can also be extended to create nanoscale protein arrays. Additionally; its adaptability for the assembly of arbitrary proteins and antibodies provides great potentials for biosensor and bio-microelectromechanical systems (MEMS) applications. © 2008 The Association for Laboratory Automation;
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页码：237 / 242
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