Dendrimer-activated surfaces for high density and high activity protein chip applications.
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.
Pathak
Srikant
S
Materials Chemistry, Sandia National Laboratories, MS 9403, 7011 East Avenue, Livermore, California 94550, USA. spathak@sandia.gov
Singh
Anup K
AK
McElhanon
James R
JR
Dentinger
Paul M
PM
D000469:Alkaline Phosphatase / Q000737:chemistry; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D050091:Dendrimers / Q000737:chemistry*; D007093:Imidazoles / Q000737:chemistry*; D055767:Immobilized Proteins; D008958:Models, Molecular; D015394:Molecular Structure; D011126:Polypropylenes / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D012821:Silanes / Q000737:chemistry*; D013499:Surface Properties
D050091:Dendrimers; D007093:Imidazoles; D055767:Immobilized Proteins; D011126:Polypropylenes; D011506:Proteins; D012821:Silanes; C443641:poly(propyleneimine); C082343:silanol; C006900:N,N-carbonyldiimidazole; D000469:Alkaline Phosphatase