[Proteomics Clinical Applications] The application of a microfluidic reactor including spontaneously
In this study the application of a newly developed microfluidic immobilized enzymatic reactor (IMER) designed to accelerate protein digestion in clinical samples is presented. The IMER contains trypsin adsorbed on the porous surface of a PDMS microfluidic chip. Human tear with its relatively low volume and high protein content was collected and used for testing the digestion efficiency of the IMER. With the use of CZE peptide mapping, the efficiency and reproducibility of the reactor were investigated. No significant difference was observed in the CZE peptide profiles of the same tear sample digested in-solution or via microfluidic IMER. LC-MS measurements showed that the microfluidic IMER digestion enabled the identification of more proteins compared to standard in-solution digestion and those proteins that were identified with both digestion methods had higher sequence coverage when digested with the IMER. The proposed reactor is well-suited for rapid and efficient protein digestion and even 8 digestions can be carried out simultaneously. The PDMS chip is inexpensive and easy to fabricate, thus its application can be an attractive alternative for proteomic related research.
Adam Kecskemeti, Cynthia Nora Nagy, Eva Csosz, Gergo Kallo, Attila Gaspar Accepted manuscript online: 8 July 2017Full publication history DOI: 10.1002/prca.201700055