Patterning of functional antibodies and other proteins by photolithography of silane monolayers.
We have demonstrated the assembly of two-dimensional patterns of functional antibodies on a surface. In particular, we have selectively adsorbed micrometer-scale regions of biotinylated immunoglobulin that exhibit specific antigen binding after adsorption. The advantage of this technique is its potential adaptability to adsorbing arbitrary proteins in tightly packed monolayers while retaining functionality. The procedure begins with the formation of a self-assembled monolayer of n-octadecyltrimethoxysilane (OTMS) on a silicon dioxide surface. This monolayer can then be selectively removed by UV photolithography. Under appropriate solution conditions, the OTMS regions will adsorb a monolayer of bovine serum albumin (BSA), while the silicon dioxide regions where the OTMS has been removed by UV light will adsorb less than 2% of a monolayer, thus creating high contrast patterned adsorption of BSA. The attachment of the molecule biotin to the BSA allows the pattern to be replicated in a layer of streptavidin, which bonds to the biotinylated BSA and in turn will bond an additional layer of an arbitrary biotinylated protein. In our test case, functionality of the biotinylated goat antibodies raised against mouse immunoglobulin was demonstrated by the specific binding of fluorescently labeled mouse IgG.
Mooney
J F
JF
Department of Physics, University of Colorado, Boulder 80309-0390, USA.
Hunt
A J
AJ
McIntosh
J R
JR
Liberko
C A
CA
Walba
D M
DM
Rogers
C T
CT
D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D018397:Equidae; D006041:Goats; D051379:Mice; D008856:Microscopy, Fluorescence; D017646:Organosilicon Compounds; D011487:Protein Conformation; D012821:Silanes; D012822:Silicon Dioxide
D000906:Antibodies; D017646:Organosilicon Compounds; D012821:Silanes; C102207:n-octadecyltrimethoxysilane; D012822:Silicon Dioxide