Rapid enrichment of target peptide by using microfluidic chip for mass spectrometry detection

• Main Authors: Wei Ting Su, 蘇瑋婷
• Other Authors: Y. H. Lin
• Format: Others
• Language: zh-TW
• Published: 2016
• Online Access: http://ndltd.ncl.edu.tw/handle/7x4euc

碩士 === 長庚大學 === 醫療機電工程研究所 === 104 === Sample pretreatment is important for mass spectrometry assay. Many protein related researches adopt KingFisherTM (Thermo Scientific) for the automatic peptide enrichment. However, purification of target peptide in KingFisherTM takes user more than 2 hours. In this study, we developed a microfluidic chip that can be used to rapidly enrich peptide sample for mass spectrometry assay within 30 min. The chip is with dimensions of 40 mm × 90 mm and consists of two polydimethylsiloxane structure layers and one thin polydimethylsiloxane layers. Five reagents reservoirs, five micro-mixer and four micro-valves were designed in a single chip. Peptide immunoprecipitation, wash, and peptide elution procedures can be automatically conducted in the microfluidic chip. Furthermore, with aid of pneumatic micro-mixer operating with 5 Hz and -75 kPa, the peptide immunoprecipitation process can be done within 10 minutes and the capture recovery was estimated to be ~33%, which is about 80% of recovery compared to KingFisherTM under standard operating time. As a whole, this microfluidic system may provide a powerful platform for the rapid pretreatment of peptide sample for MALDI-TOF mass spectrometry assay.