Dendrimer-Activated Surfaces for High Density and High Activity Protein Chip Applications

Abstract

Highly functional Si and glass surfaces for protein immobilization have been prepared by a facile activation of native surface silanol groups. Poly(propyleneimine) dendrimers of generations 1-5 were immobilized onto the surface using a facile room-temperature coupling procedure that involved activation of native silanol groups of glass using 1,1'-carbonyldiimidazole under anhydrous conditions. The dendrimer-coated surfaces were used to immobilize proteins and were characterized with respect to surface loading and activity. A number of different chemical, physical, and biochemical techniques including contact angle measurement, ellipsometry, and fluorescence microscopy were used to characterize the resulting surfaces. Increasing the dendrimer generation past G-3 led to increased surface amine content, immobilized protein concentration, and the activity of immobilized alkaline phosphatase (used as a test system). Very high activity of the immobilized proteins in the case of higher generation (G-4 and G-5) dendrimers led us to conclude that such an approach has true potential for creating highly functional surfaces for protein chip applications.

Fabrication of Robust Biomolecular Patterns by Reactive Microcontact Printing onN-Hydroxysuccinimide Ester-Containing Polymer Films

“…However, the optimum coverage or activity of molecules immobilized in patterns through mere physical interactions are often difficult to achieve because they can be washed away during processing. [11] Covalent linkages, on the other hand, are thought to provide more specific and stronger attachment. [12,13] In general, covalent attachment utilizes bifunctional linker chemistry that involves, e.g, amide-or imine-bond formation, gold-thiol interactions, or silane chemistry.…”

Dendrimer porphyrin-based self-assembled nano-devices for biomedical applications

“…DP-coated surfaces can load more proteins and more protein activity than planar surfaces. [64][65][66][67] Furthermore, we determined the relative amounts of dendrimer immobilized on the surface and monitored the enzymecatalyzed reactions using fluorescence microscopy by means of the fluorescent properties of the DP. 66 To prepare a DP-coated surface, the surface of a silicon wafer was covered with positively charged amine groups using 3-aminopropyltriethoxysilane deposited DP via electrostatic interaction in a silanization reaction (Figure 12).…”

Spatially addressable protein array: ssDNA‐directed assembly for antibody microarray

“…The choice of a carboxyl dendrimer is critical but not demonstrated in early attempts to fabricate a high-density and low nonspecific antibody array [21][22][23]. We have explored the use of such amine dendrimer-modified surfaces but high nonspecific adsorption of cellular biomolecules rendered this approach unacceptable.…”