Lithographic techniques and surface chemistries for the fabrication of PEG-passivated protein microarrays.
This article presents a new technique to fabricate patterns of functional molecules surrounded by a coating of the inert poly(ethylene glycol) (PEG) on glass slides for applications in protein microarray technology. The chief advantages of this technique are that it is based entirely on standard lithography processes, makes use of glass slides employing surface chemistries that are standard in the microarray community, and has the potential to massively scale up the density of microarray spots. It is shown that proteins and antibodies can be made to self-assemble on the functional patterns in a microarray format, with the PEG coating acting as an effective passivating agent to prevent non-specific protein adsorption. Various standard surface chemistries such as aldehyde, epoxy and amine are explored for the functional layer, and it is conclusively demonstrated that only an amine-terminated surface satisfies all the process constraints imposed by the lithography process sequence. The effectiveness of this microarray technology is demonstrated by patterning fluorescent streptavidin and a fluorescent secondary antibody using the well-known and highly specific interaction between biotin and streptavidin.
Kannan
Balaji
B
Department of Mechanical Engineering, University of California, Berkeley, CA 94720, USA. balajik@me.berkeley.edu
Castelino
Kenneth
K
Chen
Fanqing Frank
FF
Majumdar
Arun
A
D000327:Adsorption; D001710:Biotin; D005456:Fluorescent Dyes; D005464:Fluorobenzenes; D011092:Polyethylene Glycols; D040081:Protein Array Analysis; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D019809:Streptavidin
C513915:AlexaFluor 488 carboxylic acid tetrafluorophenyl ester; D005456:Fluorescent Dyes; D005464:Fluorobenzenes; D011506:Proteins; D011092:Polyethylene Glycols; D001710:Biotin; D019809:Streptavidin