Adsorption of avidin on microfabricated surfaces for protein biochip applications.
The adsorption of the protein avidin from hen egg white on patterns of silicon dioxide and platinum surfaces on a microchip and the use of fluorescent microscopy to detect binding of biotin are described. A silicon dioxide microchip was formed using plasma-enhanced chemical vapor deposition while platinum was deposited using radiofrequency sputtering. After cleaning using a plasma arc, the chips were placed into solutions containing avidin or bovine serum albumin. The avidin was adsorbed onto the microchips from phosphate-buffered saline (PBS) or from PBS to which ammonium sulfate had been added. Avidin was also adsorbed onto bovine serum albumin (BSA)-coated surfaces of oxide and platinum. Fluorescence microscopy was used to confirm adsorption of labeled protein, or the binding of fluorescently labeled biotin onto previously adsorbed, unlabeled avidin. When labeled biotin in PBS was presented to avidin adsorbed onto a BSA-coated microchip, the fluorescence signal was significantly higher than for avidin adsorbed onto the biochip alone. The results show that a simple, low-cost adsorption process can deposit active protein onto a chip in an approach that has potential application in the development of protein biochips for the detection of biological species.
Bashir
R
R
School of Electrical and Computer Engineering, 1285 EE Building, Purdue University, West Lafayette, Indiana 47907, USA. bashir@ecn.purdue.edu
Gomez
R
R
Sarikaya
A
A
Ladisch
M R
MR
Sturgis
J
J
Robinson
J P
JP
D000327:Adsorption; D000818:Animals; D001360:Avidin / Q000737:chemistry*; D015374:Biosensing Techniques; D001710:Biotin / Q000737:chemistry; D002645:Chickens; D008856:Microscopy, Fluorescence; D010047:Ovalbumin / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry
D011506:Proteins; D001360:Avidin; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D001710:Biotin; D012822:Silicon Dioxide; D010047:Ovalbumin