Effective enhancement of fluorescence detection efficiency in protein microarray assays: application of a highly fluorinated organosilane as the blocking agent on the background surface by a facile vapor-phase deposition process.
Protein microarrays are emerging as an important enabling technology for the simultaneous investigation of complicated interactions among thousands of proteins. The solution-based blocking protocols commonly used in protein microarray assays often cause cross-contamination among probes and diminution of protein binding efficiency because of the spreading of blocking solution and the obstruction formed by the blocking molecules. In this paper, an alternative blocking process for protein microarray assays is proposed to obtain better performance by employing a vapor-phase deposition method to form self-assembled surface coatings using a highly fluorinated organosilane as the blocking agent on the background surfaces. Compared to conventional solution-based blocking processes, our experimental results showed that this vapor-phase process could shorten the blocking time from hours to less than 10 min, enhance the binding efficiency by up to 6 times, reduce the background noise by up to 16 times, and improve the S/N ratio by up to 64 times. This facile blocking process is compatible with current microarray assays using silica-based substrates and can be performed on many types of silane-modified surfaces.
Hsieh
Hsin-Yi
HY
Institute of NanoEngineering and MicroSystems (NEMS), National Tsing Hua University, Hsinchu 30013, Taiwan R.O.C.
Wang
Pen-Cheng
PC
Wu
Chun-Lung
CL
Huang
Chi-Wen
CW
Chieng
Ching-Chang
CC
Tseng
Fan-Gang
FG
D005453:Fluorescence; D005461:Fluorine / Q000737:chemistry*; D005740:Gases / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D012821:Silanes / Q000737:chemistry*; D013997:Time Factors; D014835:Volatilization
D005740:Gases; D012821:Silanes; D005461:Fluorine