Target peptide enrichment microfluidic chip for rapid detection of oral squamous cell carcinoma using stable isotope standards and capture by anti-peptide antibodies

Yen Heng Lin*, Chia Chun Wu, Wei Ting Su, Pei Chi Tseng, Ya Yu Hsueh, Yung Chin Hsiao, Kai Ping Chang, Jau Song Yu*, Yun Ju Chuang

*此作品的通信作者

研究成果: 期刊稿件文章同行評審

5 引文 斯高帕斯(Scopus)

摘要

The use of antibodies against surrogate peptides for target protein enrichment combined with stable isotope standards is one of the important steps in the quantitative analysis of protein samples via mass spectrometry. Currently, commercially available automated instruments such as KingFisher™ are used to perform immunoprecipitation, wash, and elution of protein samples using magnetic beads and immune reactions. However, the use of these instruments for protein or peptide purification takes over 2 h. Moreover, numerous samples (e.g., 96) need to be processed at once to meet the operating cost, and the consumables used in conjunction with them are expensive. Thus, this study developed an automated microfluidic chip that could rapidly purify target peptides in saliva samples within 50 min. The chip is made of five connected pneumatic micromixers that automate the immunoprecipitation of target peptide from the samples. Under optimum operating conditions of 5 Hz and −75 kPa, the recovery rate of peptides achieved by the micromixer in an operating time of 30 min was similar to that achieved by KingFisher™ in the standard operating time of 2 h. The chip-processed samples could be used for the quantitative detection of the oral cancer biomarker protein MMP1 via mass spectrometry with a detection limit of 6.75 ng/mL. Six saliva samples were used to verify the feasibility of the chip for clinical sample use. The MMP1 concentration measured using our sample treatment method was comparable to that treated using a commercially available instrument.

原文英語
文章編號128607
期刊Sensors and Actuators, B: Chemical
322
DOIs
出版狀態已出版 - 01 11 2020

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Publisher Copyright:
© 2020 Elsevier B.V.

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