Protein patterning on silicon-based surface using background hydrophobic thin film.
A new and convenient protein patterning method on silicon-based surface was developed for protein array by spin coating of hydrophobic thin film (CYTOP). Photolithographic lift-off process was used to display two-dimensional patterns of spatially hydrophilic region. The background hydrophobic thin film was used to suppress nonspecific protein binding, and the hydrophilic target protein binding region was chemically modified to introduce aldehyde group after removal of the photoresist layer. The difference in surface energy between the hydrophilic pattern and background hydrophobic film would induce easier covalent binding of proteins onto defined hydrophilic areas having physical and chemical constraints. Below 1 microg/ml of total protein concentration, the CYTOP hydrophobic film effectively suppressed nonspecific binding of the protein. During the process of protein patterning, inherent property of the hydrophobic thin film was not changed judging from static and dynamic contact angle survey. Quantitative analysis of the protein binding was demonstrated by streptavidin-biotin system.
Lee
Chang-Soo
CS
School of Chemical Engineering, Seoul National University, Kwanak-Ku, Seoul 151-742, South Korea.
Lee
Sang-Ho
SH
Park
Sung-Soo
SS
Kim
Yong-Kweon
YK
Kim
Byung-Gee
BG
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D001710:Biotin / Q000032:analysis* / Q000737:chemistry; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis*; D005240:Feasibility Studies; D057927:Hydrophobic and Hydrophilic Interactions; D008567:Membranes, Artificial; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D011485:Protein Binding; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012825:Silicon / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods; D019809:Streptavidin / Q000737:chemistry*
D020099:Coated Materials, Biocompatible; D008567:Membranes, Artificial; D001710:Biotin; D019809:Streptavidin; D012825:Silicon